Methods and compositions for treating rheumatoid arthritis

ABSTRACT

This application relates to methods and compositions for treating rheumatoid arthritis by administering a combination therapy comprising methotrexate and an antibody to alpha-4 integrin or an immunologically active antigen binding fragment in therapeutically effective amounts. The application also relates generally to methods and compositions for treating rheumatoid arthritis by administering a combination therapy comprising methotrexate and small molecule alpha-4 integrin antagonist that inhibits the alpha-4 integrin (α4 integrin) interaction with VCAM-1. The invention further relates to methods of preparing the compounds and methods of using the compounds and compositions.

FIELD OF THE INVENTION

This application relates to methods and compositions for treatingrheumatoid arthritis by administering a combination therapy comprisingmethotrexate and an antibody to alpha-4 integrin or an immunologicallyactive antigen binding fragment in therapeutically effective amounts.The application also relates generally to methods and compositions fortreating rheumatoid arthritis by administering a combination therapycomprising methotrexate and small molecule alpha-4 integrin antagonistthat inhibits the alpha-4 integrin (α4 integrin) interaction withVCAM-1. The invention further relates to methods of preparing thecompounds and methods of using the compounds and compositions.

BACKGROUND OF THE INVENTION

Inflammation

Inflammation is a response of vascularized tissues to infection orinjury and is affected by adhesion of leukocytes to the endothelialcells of blood vessels and their infiltration into the surroundingtissues. In normal inflammation, infiltrating leukocytes release toxicmediators to kill invading organisms, phagocytize debris and dead cells,and play a role in tissue repair and the immune response. However, inpathologic inflammation, infiltrating leukocytes are over-responsive andcan cause serious or fatal damage. See, e.g., Hickey,Psychoneuroimmunology II (Academic Press 1990).

The integrins are a family of cell-surface glycoproteins involved incell-adhesion, immune cell migration and activation. Alpha-4 (α4)integrin is expressed by all circulating leukocytes except neutrophils,and forms heterodimeric receptors in conjunction with either the beta1(β1) or beta7 (β7) integrin subunits. Both alpha-4 beta-1 (α4β1) andalpha-4 beta-7 (α4β7) play a role in the migration of leukocytes acrossthe vascular endothelium (Springer et al., Cell, 1994 76: 301-14;Butcher et al., Science, 1996, 272: 60-6) and contribute to cellactivation and survival within the parenchyma (Damle et al., J.Immunol., 1993; 151: 2368-79; Koopman et al., J. Immunol., 1994, 152:3760-7; Leussink et al., Acta Neuropathol., 2002, 103: 131-136). α4β1 isconstitutively expressed on lymphocytes, monocytes, macrophages, mastcells, basophils and eosinophils.

α4β1 (also known as very late antigen-4, VLA-4), binds to vascular celladhesion molecule-1 (VCAM-1) (Lobb et al., J. Clin. Invest., 1994, 94:1722-8), which is expressed by the vascular endothelium at many sites ofchronic inflammation (Bevilacqua et al., 1993 Annu. Rev. Immunol., 11:767-804; Postigo et al., 1993 Res. Immunol.,144: 723-35). α4β1 has otherligands, including fibronectin and other extracellular matrix (ECM)components.

Intercellular adhesion mediated by α4β1 and other cell surface receptorsis associated with a number of inflammatory responses. At the site of aninjury or other inflammatory stimulus, activated vascular endothelialcells express molecules that are adhesive for leukocytes. The mechanicsof leukocyte adhesion to endothelial cells involves, in part, therecognition and binding of cell surface receptors on leukocytes to thecorresponding cell surface molecules on endothelial cells. Once bound,the leukocytes migrate across the blood vessel wall to enter the injuredsite and release chemical mediators to combat infection.

Rheumatoid Arthritis

Rheumatoid arthritis (“RA”) is a chronic inflammatory disease thatcauses pain, swelling, stiffness, and loss of function, primarily thejoints. RA is estimated to affect approximately 1 percent of the world'spopulation. In the U.S. alone, an estimated 2.1 million people sufferfrom the disease. This relatively high frequency suggests a complexetiology and pathogenesis.

The disease process leading to RA begins in the synovium, the membranethat surrounds a joint creating a protective sac. In healthyindividuals, the synovium produces synovial fluid that lubricates,nourishes and protects joint tissues. This clear fluid lubricates andnourishes the cartilage and bones inside the joint capsule. Inindividuals suffering from RA, the immune system, for unknown reasons,attacks the cells inside synovium. Leukocytes infiltrate from thecirculation into the synovium causing continuous abnormal inflammation(i.e., synovitis). Consequently, the synovium becomes inflamed, causingwarmth, redness, swelling, and pain. The collagen in the cartilage isgradually destroyed, narrowing the joint space and eventually damagingbone. The inflammation causes erosive bone damage in the affected area.During this process, the cells of the synovium grow and divideabnormally, making the normally thin synovium thick and resulting in ajoint that is swollen and puffy to the touch. See, e.g., Paul,Immunology (3d ed., Raven Press, 1993).

It is believed that bone damage begins during the first year or two thata person has the disease. This is one reason why early diagnosis andtreatment are important in the management of RA. As the diseaseprogresses, abnormal synovial cells begin to invade and destroy thecartilage and bone within the joint. The surrounding muscles, ligaments,and tendons that support and stabilize the joint become weak and unableto work normally. RA also causes more generalized bone loss that maylead to osteoporosis, making bones fragile and more prone to fracture.All of these effects cause the pain, impairment and deformitiesassociated with RA.

Although RA almost always develops in the wrists and knuckes, somepatients experience the effects of the disease in places other than thejoints. For instance, the knees and the ball of the foot are oftenaffected as well. Often, many joints may be involved, and even the spinecan be affected. In about 25% of people with RA, inflammation of smallblood vessels can cause rheumatoid nodules, or lumps, under the skin.These are bumps under the skin that often form close to the joints. Asthe disease progresses, fluid may also accumulate, particularly in theankles. Many patients with RA also develop anemia, or a decrease in thenormal number of red blood cells. Other less prevalent effects includeneck pain, dry eyes and dry mouth. On rare occasions, patients may alsodevelop inflammation of the blood vessels, the lining of the lungs, orthe sac enclosing the heart.

RA has several special features that differentiate it from other typesof arthritis. For example, RA generally occurs in a symmetricalpattern—if one knee or hand is involved, the other one is also. Thedisease often affects the wrist joints and the fingerjoints closest tothe hand. RA usually first affects the small joints of the hands andfeet, but may also involve the wrists, elbows, ankles and knees. It canalso affect other parts of the body besides the joints. In addition,patients with the disease may have fatigue, occasional fever, and ageneral sense of not feeling well (malaise).

Another distinct feature of RA is the variance between individuals. Forsome, it lasts only a few months or a year or two and subsides withoutcausing any noticeable damage. Other people have mild or moderatedisease, with periods of worsening symptoms (flares) and periods inwhich they feel better (remissions). In severe cases, the disease ischronically active most of the time, lasting for many years, and leadingto serious joint damage and disability.

RA encompasses a number of disease subtypes, such as Felty's syndrome,seronegative RA, “classical” RA, progressive and/or relapsing RA, and RAwith vasculitis. Some experts classify the disease into type 1 or type2. Type 1, the less common form, lasts a few months at most and leavesno permanent disability. Type 2 is chronic and lasts for years,sometimes for life.

RA is believed to be one of several “autoimmune” diseases (“auto” meansself), so-called because a person's immune system attacks his or her ownbody tissues. Although much has been learned about the process leadingto RA, researchers have yet to uncover all of the factors that lead tothis disease. One prevalent theory is that a combination of factorstrigger RA, including an abnormal autoimmune response, geneticsusceptibility, environmental, biologic factors, hormonal, andreproductive factors. Nonetheless, despite intensive research, the causeof RA remains obscure. See El-Gabalawy et al., ARTHRITIS RES. 4(suppl3):S297-S301 (2002).

RA occurs across all races and ethnic groups. Although the disease oftenbegins in middle age and occurs with increased frequency in olderpeople, children and young adults may also develop juvenile RA. Likeother forms of arthritis, RA exhibits a clear gender bias: approximatleytwo to three times as many women as men have the disease (Lawrence etal., Arthritis Rheum., 1998, 41:778-799). However, a geneticpredisposition has been identified and, in white populations, localizedto a pentapeptide in the HLA-DR 1 locus of class II histocompatibilitygenes. Environmental factors may also play a role. Immunologic changesmay be initiated by multiple factors.

Prominent immunologic abnormalities that may be important inpathogenesis include immune complexes found in joint fluid cells and invasculitis. Plasma cells produce antibodies that contribute to thesecomplexes. Lymphocytes that infiltrate the synovial tissue are primarilyT helper cells, which can produce pro-inflammatory cytokines. Increasedadhesion molecules contribute to inflammatory cell emigration andretention in the synovial tissue.

The onset is usually insidious, with progressive joint involvement, butmay be abrupt, with simultaneous inflammation in multiple joints.Tenderness in nearly all inflamed joints and synovial thickening arecommon. Initial manifestations may occur in any joint.

Stiffness lasting less than 30 minutes on arising in the morning orafter prolonged inactivity is common. Subcutaneous rheumatoid nodulesare not usually an early manifestation. Visceral nodules, vasculitiscausing leg ulcers or mononeuritis multiplex, pleural or pericardialeffusions, lymphadenopathy, Felty's syndrome, Sjögren's syndrome, andepiscleritis are other manifestations. As many as 75% of patientsimprove symptomatically with conservative treatment during the firstyear of disease. However, less than 10% are eventually severely disableddespite full treatment. The disease greatly affects the lives of most RApatients. Complete bed rest is occasionally indicated for a short periodduring the most active, painful stage of severe disease. In less severecases, regular rest should be prescribed.

Nonsteroidal anti-inflammatory drugs may provide important symptomaticrelief and may be adequate as simple therapy for mild RA, but they donot appear to alter the long-term course of disease. Salicylates, suchas aspirin, may be used for treatment.

Gold compounds usually are given in addition to salicylates or otherNSAIDs if the latter do not sufficiently relieve pain or suppress activejoint inflammation. In some patients, gold may produce clinicalremission and decrease the formation of new bony erosions. Parenteralpreparations include gold sodium thiomalate or gold thioglucose. Goldshould be discontinued when any of the above manifestations appear.Minor toxic manifestations (e.g., mild pruritus, minor rash) may beeliminated by temporarily withholding gold therapy, then resuming itcautiously about 2 wk after symptoms have subsided. However, if toxicsymptoms progress, gold should be withheld and the patient given acorticosteroid. A topical corticosteroid or oral prednisone 15 to 20mg/day in divided doses is given for mild gold dermatitis; larger dosesmay be needed for hematologic complications. A gold chelating drug,dimercaprol 2.5 mg/kg IM, may be given up to four to six times/day forthe first 2 days and bid for 5 to 7 days after a severe gold reaction.

Hydroxychloroquine can also control symptoms of mild or moderatelyactive RA. Toxic effects usually are mild and include dermatitis,myopathy, and generally reversible corneal opacity. However,irreversible retinal degeneration has been reported. Sulfasalazine mayalso be used for treatment of RA.

Oral penicillamine may have a benefit similar to gold and may be used insome cases if gold fails or produces toxicity in patients with activeRA. Side effects requiring discontinuation are more common than withgold and include marrow suppression, proteinuria, nephrosis, otherserious toxic effects (eg, myasthenia gravis, pemphigus, Goodpasture'ssyndrome, polymyositis, a lupuslike syndrome), rash, and a foul taste.

Steroids are the most effective short-term anti-inflammatory drugs.However, their clinical benefit for RA often diminishes with time.Steroids do not predictably prevent the progression of jointdestruction. Furthermore, severe rebound follows the withdrawal ofcorticosteroids in active disease. Contraindications to steroid useinclude peptic ulcer, hypertension, untreated infections, diabetesmellitus, and glaucoma.

Immunosuppressive drugs are increasingly used in management of severe,active RA. However, major side effects can occur, including liverdisease, pneumonitis, bone marrow suppression, and, after long-term useof azathioprine and malignancy.

Whatever may be the actual cause, there is no cure for RA, and althoughthe disease is not fatal, disease complications and symptoms may persistthroughout an individual's lifetime, and may even shorten survival by afew years. Affected joints may become deformed, and the performance ofeven ordinary tasks may be very difficult or impossible.

SUMMARY OF THE INVENTION

Based on the above, new compositions and methods of preventingrheumatoid arthritis and treating the symptoms of rheumatoid arthritisare needed such that patients can have better quality of life.

The invention relates to combination therapies comprising methotrexateand an antibody to alpha-4 integrin or an immunologically active antigenbinding fragment thereof or a small molecule alpha-4 integrin antagonistfor use in a subject in need thereof. Preferably, the subject is amammal. More preferably, the mammal is human.

The invention also relates to methods of treating rheumatoid arthritisin a subject in need thereof comprising administering in therapeuticallyeffective amounts, a combination therapy comprising methotrexate and anantibody to alpha-4 integrin or an immunologically active antigenbinding fragment thereof or a small molecule alpha-4 integrinantagonist. Preferably, the subject is a mammal. More preferably, themammal is human.

The invention further relates to regimes for the treatment of rheumatoidarthritis which comprises administering to a subject in need thereofabout 2 mg to about 20 mg of methotrexate and a therapeuticallyeffective amount of a methotrexate and an antibody to alpha-4 integrinor an immunologically active antigen binding fragment thereof or a smallmolecule alpha-4 integrin antagonist. Preferably, the subject is amammal. More preferably, the mammal is human.

In yet another embodiment, the invention relates to the use of thecombination therapies as described herein in the preparation of amedicament for the treatment of rheumatoid arthritis.

These and other objects, advantages, and features of the invention willbecome apparent to those persons skilled in the art upon reading thedetails of the methods and formulations as more fully described below.

BRIEF DESCRIPTIONS OF THE DRAWINGS

FIG. 1. FIG. 1 shows the effect of anti-alpha 4-integrin antibody invivo prophylactically (upper panel), semi-therapeutic dosing (middlepanel) and therapeutic dosing (lower panel).

FIG. 2. FIG. 2 shows the effect of anti-alpha4 antibodies (upper panel),anti-α4β7 (LPAM-1) antibodies (middle panel) and anti-VACM-1 antibodiesin vivo using CIA models (lower panel).

FIG. 3. FIG. 3 shows the effects of anti-VLA-4, anti-VCAM-2 andanti-LPAM-1 antibodies in vivo in CIA models at semi-therapeutic dosing.

FIG. 4. FIG. 4 shows the effect of anti-VLA-4 antibody in vivo in CIAmodel at therapeutic dosing (upper panel). Effect of the compound ofFormula P in vivo in CIA model at therapeutic dosing (lower panel).

FIG. 5. FIG. 5 shows the valuation of compounds in AIA Animal Model.

FIG. 6. FIG. 6 shows comparative effects of anti-alpha 4 antibodies andthe compounds of Formulae W and Y in rat CIA model.

FIG. 7. FIG. 7 shows prophylactic treatment with anti-alpha 4 antibodies(PS/2) in the CIA Animal Model.

FIG. 8. FIG. 8 shows the therapeutic treatment with anti-alpha 4antibody (GG5/3) in the AIA Animal Model.

FIG. 9. FIG. 9 shows the therapeutic treatment with anti-alpha 4antibody (GG5/3) in the AIA Animal Model.

FIG. 10. FIG. 10 shows the results of dosage regimen for therapeutictreatment with small molecule alpha-4 antagonists in the AIA AnimalModel.

FIG. 11. FIG. 11 shows the potency and specificity of compounds in theAIA Animal Model.

DETAILED DESCRIPTION OF THE INVENTION

In accordance with this detailed description, the followingabbreviations and definitions apply. It must be noted that as usedherein, the singular forms “a”, “an”, and “the” include plural referentsunless the context clearly dictates otherwise. Thus, for example,reference to “an antibody” includes a plurality of such antibodies andreference to “the dosage” includes reference to one or more dosages andequivalents thereof known to those skilled in the art, and so forth.

The publications discussed herein are provided solely for theirdisclosure prior to the filing date of the present application. Nothingherein is to be construed as an admission that the present invention isnot entitled to antedate such publication by virtue of prior invention.Further, the dates of publication provided may be different from theactual publication dates, which may need to be independently confirmed.

Where a range of values is provided, it is understood that eachintervening value, to the tenth of the unit of the lower limit unlessthe context clearly dictates otherwise, between the upper and lowerlimit of that range and any other stated or intervening value in thatstated range is encompassed within the invention. The upper and lowerlimits of these smaller ranges may independently be included in thesmaller, subject to any specifically excluded limit in the stated range.Where the stated range includes one or both of the limits, rangesexcluding either both of those included limits are also included in theinvention. Also contemplated are any values that fall within the citedranges.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can also beused in the practice or testing of the present invention, the preferredmethods and materials are now described. All publications mentionedherein are incorporated herein by reference to disclose and describe themethods and/or materials in connection with which the publications arecited.

DEFINITIONS

By “protein” is meant to include but is not limited to immunoglobulins,enzymes, receptor, and fragments thereof. Although discussion of theformulation is provided in reference to an antibody or immunoglobulin,other proteins are contemplated as interchangeable in the formulationsdisclosed.

By “immunoglobulin” is meant to include, but is not limited to, anantibody and antibody fragment (such as scFv, Fab, Fc, F(ab′)₂), andother genetically engineered portions of antibodies. Depending on theamino acid sequence of the constant domain of their heavy chains,immunoglobulins can be assigned to different classes. There are fivemajor classes of immunoglobulins: IgA, IgD, IgE, IgG and IgM. Several ofthese may be further divided into subclasses (isotypes), e.g., IgG-1,IgG-2, IgG-3, and IgG-4; IgA-1 and IgA-2. The heavy chain constantdomains that correspond to the different classes of immunoglobulins arecalled alpha (α), delta (Δ), epsilon (ε), gamma (γ), and mu (μ),respectively. The subunit structures and three-dimensionalconfigurations of different classes of immunoglobulins are well known.Preferably, the immunoglobulin recognizes and binds to alpha-4 integrin.

The term “antibody” is used in the broadest sense and includesmonoclonal antibodies (including agonist and antagonist antibodies),antibody compositions with polyepitopic specificity, and antibodyfragments (e.g., Fab, F(ab′)₂, scFv and Fv), so long as they exhibit thedesired biological activity. “Antibody” is meant to include polyclonalantibodies, monoclonal antibodies, humanized antibodies, humanantibodies, Primatized® antibodies and other antibodies produced viagenetic engineering.

The term “monoclonal antibody” as used herein refers to an antibodyobtained from a population of substantially homogeneous antibodies,i.e., the individual antibodies comprising the population are identicalexcept for possible naturally occurring mutations that may be present inminor amounts. Monoclonal antibodies are highly specific, being directedagainst a single antigenic site. Furthermore, in contrast toconventional (polyclonal) antibody preparations, which typically includedifferent antibodies directed against different determinants (epitopes),each monoclonal antibody is directed against a single determinant on theantigen. In addition to their specificity, the monoclonal antibodies areadvantageous in that they are synthesized by the hybridoma culture,uncontaminated by other immunoglobulins. The modifier “monoclonal”indicates the character of the antibody as being obtained from asubstantially homogeneous population of antibodies, and is not to beconstrued as requiring production of the antibody by any particularmethod. For example, the monoclonal antibodies to be used in accordancewith the present invention may be made by the hybridoma method firstdescribed by Kohler et al., 1975 Nature 256: 495 or by the manyimprovements thereon derived in the intervening period. Some of theseare discussed for instance in, Harlow et al., USING ANTIBODIES: ALABORATORY MANUAL—PORTABLE PROTOCOL NO. 1 (Cold Spring Harbor Press, NewYork 1998); Harlow et al., ANTIBODIES: A LABORATORY MANUAL (Cold SpringHarbor Press, New York 1988); and Shepherd et al., MONOCLONALANTIBODIES: A PRACTICAL APPROACH (Oxford University Press, 2000).

The term “monoclonal antibodies” also includes “chimeric” antibodies(immunoglobulins) in which a portion of the heavy and/or light chain isidentical with or homologous to corresponding sequences in antibodiesderived from a particular species or belonging to a particular antibodyclass or subclass, while the remainder of the chain(s) is identical withor homologous to corresponding sequences in antibodies derived fromanother species or belonging to another antibody class or subclass, aswell as fragments of such antibodies, so long as they exhibit thedesired biological activity. For example, the ability to bind to alpha-4integrin. The “monoclonal antibodies” may also be isolated from phageantibody libraries using the techniques described for example inClackson et aL, 1991 Nature 352: 624-628 and Marks et al., 1991 J. Mol.Biol., 222: 581-597.

“Humanized” forms of non-human (e.g., murine, rabbit, bovine, equine,porcine, and the like) antibodies are chimeric immunoglobulins,immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab′,F(ab′)₂ or other antigen-binding subsequences of antibodies), whichcontain minimal sequence derived from non-human immunoglobulin. For themost part, humanized antibodies are human immunoglobulins (recipientantibody) in which residues from a complementary determining region(CDR) of the recipient are replaced by residues from a CDR of anon-human species (donor antibody) such as mouse, rat or rabbit havingthe desired specificity, affinity and capacity. In some instances, Fvframework residues of the human immunoglobulin are replaced bycorresponding non-human residues. Furthermore, humanized antibody maycomprise residues which are found neither in the recipient antibody. norin the imported CDR or framework sequences. These modifications are madeto further refine and optimize antibody performance. In general, thehumanized antibody will comprise substantially all of at least one, andtypically two, variable domains, in which all or substantially all ofthe CDR regions correspond to those of a non-human immunoglobulin andall or substantially all of the FR regions are those of a humanimmunoglobulin consensus sequence. The humanized antibody optimally alsowill comprise at least a portion of an immunoglobulin constant region(Fc), typically that of a human immunoglobulin.

The expression “linear antibodies” are also included by the general term“antibody” and are a pair of tandem Fd segments(V_(H)-C_(H)1-V_(H)-C_(H)1), which form a pair of antigen bindingregions. Linear antibodies can be bispecific or monospecific.

A “variant antibody” (also included by the generic term “antibody”) is amolecule which differs in amino acid sequence from a “parent” antibody'samino acid sequence by virtue of addition, deletion and/or substitutionof one or more amino acid residue(s) in the parent antibody sequence. Inthe preferred embodiment, the variant comprises one or more amino acidsubstitution(s) in one or more hypervariable region(s) of the parentantibody. For example, the variant may comprise at least onesubstitution, e.g., from about one to about ten, and preferably fromabout two to about five, in one or more hypervariable regions of theparent antibody. Ordinarily, the variant will have an amino acidsequence having at least 75% amino acid sequence identity with theparent antibody heavy or light chain variable domain sequences, morepreferably at least 80%, more preferably at least 85%, more preferablyat least 90%, and most preferably at least 95%. Identity or homologywith respect to this sequence is defined herein as the percentage ofamino acid residues in the candidate sequence that are identical withthe parent antibodyresidues, after aligning the sequences andintroducing gaps, if necessary, to achieve the maximum percent sequenceidentity. None of the N-terminal, C-terminal, or internal extensions,deletions, or insertions into the antibody sequence should be construedas affecting sequence identity or homology.

To analyze such properties, one should compare a Fab form of the variantto a Fab form of the parent antibody or a full length form of thevariant to a full length form of the parent antibody, for example,because it has been found that the format of the antibody, impacts itsactivity in the biological activity assays disclosed herein. The variantantibody of particular interest is one which displays at least about 10fold, preferably at least about 20 fold, and most preferably at leastabout 50 fold, enhancement in biological activity when compared to theparent antibody. The “parent” antibody is one which is encoded by anamino acid sequence used for the preparation of the variant. Preferably,the parent antibody has a human framework region and has human antibodyconstant region(s). For example, the parent antibody may be a humanizedor a human antibody.

An “isolated antibody” is one which has been identified and separatedand/or recovered from a component of its natural environment.Contaminant components of its natural environment are materials whichwould interfere with diagnostic or therapeutic uses for the antibody,and may include enzymes, hormones, and other proteinaceous ornonproteinaceous solutes. In preferred embodiments, the antibody will bepurified (1) to greater than 95% by weight of antibody as determined bythe Lowry method, and most preferably more than 99% by weight, (2) to adegree sufficient to obtain at least 15 residues of N-terminal orinternal amino acid sequence by use of a spinning cup sequenator, or (3)to homogeneity by SDS-PAGE under reducing or non-reducing conditionsusing Coomassie blue or, preferably, silver stain. Isolated antibodyincludes the antibody in situ within recombinant cells since at leastone component of the antibody's natural environment will not be present.Ordinarily, however, isolated antibody will be prepared by at least onepurification step.

“Antibody fragments” comprise a portion of an intact antibody, generallythe antigen binding or variable region of the intact antibody. Examplesof antibody fragments include Fab, Fab′, F(ab′)₂, and Fv fragments;diabodies; linear antibodies; single-chain antibody molecules; andmultispecific antibodies formed from antibody fragments.

“Single-chain Fv” or “sFv” antibody fragments comprise the V_(H) andV_(L) domains of antibody, wherein these domains are present in a singlepolypeptide chain. Generally, the Fv polypeptide further comprises apolypeptide linker between the V_(H) and V_(H) domains which enables thesFv to form the desired structure for antigen binding.

The term “diabodies” refers to small antibody fragments with twoantigen-binding sites, which fragments comprise a heavy chain variabledomain (V_(H)) connected to a light chain variable domain (V_(L)) in thesame polypeptide chain (V_(H)−V_(L)). By using a linker that is tooshort to allow pairing between the two domains on the same chain, thedomains are forced to pair with the complementary domains of anotherchain and create two antigen-binding sites.

The route of antibody administration is in accord with welll knownmethods, and may include injection or infusion by intravenous,intraperitoneal, intracerebral, intramuscular, intraocular,intraarterial, or intralesional routes, or by sustained release systems.The antibody can be administered continuously by infusion or by bolusinjection. Therapeutic antibody compositions generally are placed into acontainer having a sterile access port, for example, an intravenoussolution bag or vial having a stopper pierceable by a hypodermicinjection needle.

A “stable” formulation is one in which the protein therein essentiallyretains its physical stability and/or chemical stability and/orbiological activity upon storage. Various analytical techniques formeasuring protein stability are available in the art and are reviewed inPEPTIDE AND PROTEIN DRUG DELIVERY, 247-301 (Vincent Lee ed., New York,N.Y., 1991) and Jones, 1993 Adv. Drug Delivery Rev. 10: 29-90, forexamples. Stability can be measured at a selected temperature for aselected time period as exemplified by the provided examples.

A protein, such as an antibody or fragment thereof, “retains itsphysical stability” in a pharmaceutical formulation if it shows no signsof aggregation, precipitation and/or denaturation upon visualexamination of color and/or clarity, or as measured by UV lightscattering or by size exclusion chromatography.

A protein “retains its chemical stability” in a pharmaceuticalformulation, if the chemical stability at a given time is such that theprotein is considered to still retain its biological activity. Chemicalstability can be assessed by detecting and quantifying chemicallyaltered forms of the protein. Chemical alteration may involve sizemodification (e.g., clipping), which can be evaluated using sizeexclusion chromatography, SDS-PAGE and/or matrix-assisted laserdesorption ionization/time-of-flight mass spectrometry (MALDI/TOF MS),for examples. Other types of chemical alteration include chargealteration (e.g., occurring as a result of deamidation), which can beevaluated by ion-exchange chromatography, for example.

An antibody “retains its biological activity” in a pharmaceuticalformulation, if the biological activity of the antibody at a given timeis within about 10% (within the errors of the assay) of the biologicalactivity exhibited at the time the pharmaceutical formulation wasprepared as determined in an antigen binding assay, for example.

By “isotonic” is meant that the formulation of interest has essentiallythe same osmotic pressure as human blood. Isotonic formulations willgenerally have an osmotic pressure from about 250 to 350 mOsm.Isotonicity can be measured using a vapor pressure or ice-freezing typeosmometer, for example.

As used herein, “buffer” refers to a buffered solution that resistschanges in pH by the action of its acid-base conjugate components. Thebuffer of this invention has a pH in the range from about 3.0 to about7.5; preferably from about pH 4.0 to about 7.0; more preferably fromabout pH 5.0 to about 6.5; and most preferably has a pH of about6.0±0.5. A pH of any point in between the above ranges is alsocontemplated.

A “preservative” is a compound which can be included in the formulationto essentially reduce bacterial action therein, thus facilitating theproduction of a multi-use formulation, for example. Examples ofpotential preservatives include octadecyldimethylbenzyl ammoniumchloride, hexamethonium chloride, benzalkonium chloride (a mixture ofalkylbenzyldimethylammonium chlorides in which the aikyl groups arelong-chain compounds), and benzethonium chloride. Other types ofpreservatives include aromatic alcohols such as phenol, butyl and benzylalcohol, alkyl parabens such as methyl or propyl paraben, catechol,resorcinol, cyclohexanol, 3-pentanol, and m-cresol.

By “patient” or “subject” is meant to include any mammal. A “mammal”,for purposes of treatment, refers to any animal classified as a mammal,including but not limited to humans, domestic and farm animals, and zoo,sports, or pet animals, such as dogs, horses, cats, cows, and the like.Preferably, the mammal is human.

By “Antegren™” is meant to include the antibody also known as AN100226(antibody code number) or natalizumab (USAN name). Antegren™ is arecombinant, humanized anti-alpha-4 integrin antibody. Preferably thedisease or condition being treated in the mammal is one which ismodulated.when a therapeutically effective dose of Antegren™ isadministered.

The terms “small molecule alpha-4-integrin antagonists” (i.e.,anti-alpha-4 agents and small molecule compounds) as used herein referto any agent that binds specifically to an integrin comprising analpha-4 subunit and inhibits activity of the integrin. Preferably suchmolecules bind to alpha-4 in a manner that prevents its interaction withVCAM 1 and thereby VCAM-1 signaling. This also includes agents thatspecifically bind to alpha-4 integrin as well as agents that bind to anintegrin dimer that comprises the alpha-4 integrin, e.g., alpha-4 beta-1(i.e., α4β1 integrin) or alpha-4 beta-7 (i.e., α4β7 integrin).Preferably, the agent is one that binds to alpha-4 in a manner, whichinhibits VLA4 (alpha-4) from interacting with its cognate ligand, i.e.,beta-7 or beta-1. More preferably, the anti-alpha-4 agent inhibits VLA4from interacting with VCAM-1.

The term “agent” is meant to include synthetic molecules (e.g.,antibodies, small molecules, peptides, or other synthetically producedmolecules or compounds, as well as recombinantly produced gene products)as well as naturally occurring compounds (e.g., polypeptides,antibodies, antibody fragments and the like). Preferably the agent is anantagonist of alpha-4 beta-1 integrin interaction with its cognateligand. Thus, the agent preferably binds to either VCAM-1 or to alpha-4beta-1 integrin in a manner so as to inhibit or prevent VCAM-1interaction with alpha-4 beta-1 integrin. The agent also inhibits VLA-4recruitment of immune cells inhibiting an inflammatory response, whichis responsible for the disease or condition being treated in thesubject.

The term “efficacy” as used herein in the context of a chronic dosageregime refers to the effectiveness of a particular treatment regime.Efficacy can be measured based on change the course of the disease inresponse to an agent of the present invention.

The term “success” as used herein in the context of a chronic treatmentregime refers to the effectiveness of a particular treatment regime.This includes a balance of efficacy, toxicity (e.g., side effects andpatient tolerance of a formulation or dosage unit), patient compliance,and the like. For a chronic administration regime to be considered“successful” it must balance different aspects of patient care andefficacy to produce the most favorable patient outcome.

The terms “specifically binds” or “binds specifically” as used hereinrefer to the situation in which one member of a specific binding pairwill not show any significant binding to molecules other than itsspecific binding partner (e.g., an affinity of about 1000 times or morefor its binding partner). In the present invention, the small compounds,such as N-[N-(3-pyridinesulfonyl)-L-3,3-dimethyl-4-thiaprolyl]-O-[1-methylpiperzain-4-ylcarbonyl]-L-tyrosine isopropyl ester, will not showsignificant binding to any polypeptide other than an alpha-4 integrin ora receptor comprising an alpha-4 integrin. For example, the smallcompounds used in the methods of the invention that bind to an alpha-4integrin with a binding affinity of greater than 0.3 nM are said to bindspecifically to an alpha-4 integrin.

The term “substantially similar” as used herein is intended to mean anypolypeptide that has an alteration in the sequence such that afunctionally equivalent amino acid is substituted for one or more aminoacids in the polypeptide, thus producing a change that has no orrelatively little effect on the binding properties of the polypeptide.For example, one or more amino acid residues within the sequence can besubstituted by another amino acid of a similar polarity or similar size.

The terms “elicits an immune response” and “elicits a host immuneresponse” as used herein refer to the production of an immune responseto a receptor comprising an alpha-4 integrin in a subject uponintroduction of an agent of the invention to the subject. An immuneresponse in the subject can be characterized by a serum reactivity withan alpha-4 integrin receptor that is at least twice that of an untreatedsubject, more preferably three times the reactivity of an untreatedsubject, and even more preferably at least four times the reactivity ofan untreated subject, with serum immunoreactivity measured using a serumdilution of approximately 1:100.

The terms “treating”, “treatment”, and the like are used herein to referto obtaining a desired pharmacological and physiological effect. Theeffect may be prophylactic in terms of preventing or partiallypreventing a disease, symptom or condition thereof and/or may betherapeutic in terms of a partial or complete cure of a disease,condition, symptom or adverse effect attributed to the disease. The term“treatment”, as used herein, covers any treatment of a disease in amammal, particularly a human, and includes: (a) preventing the diseasefrom occurring in a subject which may be predisposed to the disease buthas not yet been diagnosed as having it, i.e., causing the clinicalsymptoms of the disease not to develop in a subject that may bepredisposed to the disease but does not yet experience or displaysymptoms of the disease; (b) inhibiting the disease, i.e., arresting orreducing the development of the disease or its clinical symptoms; or (c)relieving the disease, i.e., causing regression of the disease and/orits symptoms or conditions. The invention is directed towards treating apatient's suffering from disease related to pathological inflammation.The present invention is involved in preventing, inhibiting, orrelieving adverse effects attributed to pathological inflammation overlong periods of time and/or are such caused by the physiologicalresponses to inappropriate inflammation present in a biological systemover long periods of time.

As used herein, “acyl” refers to the groups H—C(O)—, alkyl-C(O)—,substituted alkyl-C(O)—, alkenyl-C(O)—, substituted alkenyl-C(O)—,alkynyl-C(O)—, substituted alkynyl-C(O)— cycloalkyl-C(O)—, substitutedcycloalkyl-C(O)—, aryl-C(O)—, substituted aryl-C(O)—, heteroaryl-C(O)—,substituted heteroaryl-C(O), heterocyclic-C(O)—, and substitutedheterocyclic-C(O)— wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Acylamino” refers to the group —C(O)NRR where each R is independentlyselected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl,substituted aryl, cycloalkyl, substituted cycloalkyl, heteroaryl,substituted heteroaryl, heterocyclic, substituted heterocyclic and whereeach R is joined to form together with the nitrogen atom a heterocyclicor substituted heterocyclic ring wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Acyloxy” refers to the groups alkyl-C(O)O—, substituted alkyl-C(O)O—,alkenyl-C(O)O—, substituted alkenyl-C(O)O—, alkynyl-C(O)O—, substitutedalkynyl-C(O)O—, aryl-C(O)O—, substituted aryl-C(O)O—, cycloalkyl-C(O)O—,substituted cycloalkyl-C(O)O—, heteroaryl-C(O)O—, substitutedheteroaryl-C(O)O—, heterocyclic-C(O)O—, and substitutedheterocyclic-C(O)O— wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Alkenoxy” refers to the group “alkenyl-O—”.

“Substituted alkenoxy” refers to the group “substituted alkenyl-O—”.

“Alkenyl” refers to alkenyl group preferably having from 2 to 10 carbonatoms and more preferably 2 to 6 carbon atoms and having at least 1 andpreferably from 1-2 sites of alkenyl unsaturation.

“Lower alkenyl” refers to an alkenyl group preferably having from 2 to 6carbon atoms and having at least 1 site and preferably only 1 site ofalkenyl unsaturation (i.e., >C═C<). This term is exemplified by groupssuch as allyl, ethenyl, propenyl, butenyl, and the like.

“Substituted alkenyl” refers to alkenyl groups having from 1 to 5substituents independently selected from the group consisting of alkoxy,substituted alkoxy, acyl, acylamino, thiocarbonylamino, acyloxy, amino,amidino, alkylamidino, thioamidino, aminoacyl, aminocarbonylamino,aminothiocarbonylamino, aminocarbonyloxy, aryl, substituted aryl,aryloxy, substituted aryloxy, aryloxyaryl, substituted aryloxyaryl,halogen, hydroxyl, cyano, nitro, carboxyl, carboxylalkyl,carboxyl-substituted alkyl, carboxylcycloalkyl, carboxyl-substitutedcycloalkyl, carboxylaryl, carboxyl-substituted aryl, carboxylheteroaryl,carboxyl-substituted heteroaryl, carboxylheterocyclic,carboxyl-substituted heterocyclic, cycloalkyl, substituted cycloalkyl,guanidino, guanidinosulfone, thiol, thioalkyl, substituted thioalkyl,thioaryl, substituted thioaryl, thiocycloalkyl, substitutedthiocycloalkyl, thioheteroaryl, substituted thioheteroaryl,thioheterocyclic, substituted thioheterocyclic, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, cycloalkoxy,substituted cycloalkoxy, heteroaryloxy, substituted heteroaryloxy,hetero cyclyloxy, substituted heterocyclyloxy, oxycarbonylamino,oxythiocarbonylamino, cycloalkyloxy, substituted cycloalkyloxy,heteroaryloxy, substituted heteroaryloxy, —OS(O)₂-alkyl,—OS(O)₂-substituted alkyl, —OS(O)₂-aryl, —OS(O)₂-substituted aryl,—OS(O)₂-heteroaryl, —OS(O)₂-substituted heteroaryl,—OS(O)₂-heterocyclic, —OS(O)₂-substituted heterocyclic, —OSO₂—NRR whereR is hydrogen or alkyl, —NRS(O)₂-alkyl, —NRS(O)₂-substituted alkyl,—NRS(O)₂-aryl, —NRS(O)₂-substituted aryl, —NRS(O)₂-heteroaryl,—NRS(O)₂-substituted heteroaryl, —NRS(O)₂-heterocyclic,—NRS(O)₂-substituted heterocyclic, —NRS(O)₂—NR-alkyl,—NRS(O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl, —NRS(O)₂—NR-substitutedaryl, —NRS(O)₂—NR-heteroaryl, —NRS(O)₂—NR-substituted heteroaryl,—NRS(O)₂—NR-heterocyclic, —NRS(O)₂—NR-substituted heterocyclic where Ris hydrogen or alkyl, mono- and di-alkylamino, mono- and di-(substitutedalkyl)amino, mono- and di-arylamino, mono- and di-substituted arylamino,mono- and di-heteroarylamino, mono- and di-substituted heteroarylamino,mono- and di-heterocyclic amino, mono- and di-substituted heterocyclicamino, unsymmetric di-substituted amines having different substituentsindependently selected from the group consisting of alkyl, substitutedalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic, substituted heterocyclic and substituted alkenyl groupshaving amino groups blocked by conventional blocking groups such as Boc,Cbz, formyl, and the like or alkenyl/substituted alkenyl groupssubstituted with —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-alkenyl,—SO₂-substituted alkenyl, —SO₂-cycloalkyl, —SO₂-substituted cycloalkyl,—SO₂-aryl, —SO₂-substituted aryl, —SO₂-heteroaryl, —SO₂-substitutedheteroaryl, —SO₂-heterocyclic, —SO₂-substituted heterocyclic and —SO₂NRRwhere R is hydrogen or alkyl.

Preferably, the substituents are independently selected from the groupconsisting of alkoxy, substituted alkoxy, acyl, acylamino, acyloxy,amino, substituted amino, aminoacyl, aminocarbonylamino,aminocarbonyloxy, aryl, substituted aryl, aryloxy, substituted aryloxy,carboxyl, carboxyl esters, cyano, cycloalkyl, substituted cycloalkyl,cycloalkyloxy, substituted cycloalkyloxy, halogen, heteroaryl,substituted heteroaryl, heteroaryloxy, substituted heteroaryloxy,heterocyclic, substituted heterocyclic, hydroxyl, nitro, andoxycarbonylamino.

“Alkoxy” refers to the group “alkyl-O-” which includes, by way ofexample, methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy,sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like.

“Substituted alkoxy” refers to the group “substituted alkyl-O—”.

“Alkyl” refers to linear or branched alkyl groups preferably having from1 to 10 carbon atoms and more preferably 1 to 6 carbon atoms. This termis exemplified by groups such as methyl, t-butyl, n-heptyl, octyl andthe like.

“Lower alkyl” refers to monovalent alkyl groups having from 1 to 5carbon atoms including straight and branched chain alkyl groups. Thisterm is exemplified by groups such as methyl, ethyl, iso-propyl,n-propyl, n-butyl, iso-butyl, sec-butyl, t-butyl, n-pentyl and the like.“Lower alkyl” may be optionally substituted with a halogen, such aschloro, fluoro, bromo and the like.

“Substituted alkyl” refers to an alkyl group, of from 1 to 10 carbonatoms, having from 1 to 5 substituents independently selected from thegroup consisting of alkoxy, substituted alkoxy, acyl, acylamino,thiocarbonylamino, acyloxy, amino, amidino, alkyl amidino, thioamidino,aminoacyl, aminocarbonylamino, aminothiocarbonylamino, aminocarbonyloxy,aryl, substituted aryl, aryloxy, substituted aryloxy, aryloxylaryl,substituted aryloxyaryl, cyano, halogen, hydroxyl, nitro, carboxyl,carboxylalkyl, carboxyl-substituted alkyl, carboxyl-cycloalkyl,carboxyl-substituted cycloalkyl, carboxylaryl, carboxyl-substitutedaryl, carboxylheteroaryl, carboxyl-substituted heteroaryl,carboxylheterocyclic, carboxyl-substituted heterocyclic, cycloalkyl,substituted cycloalkyl, guanidino, guanidinosulfone, thiol, thioalkyl,substituted thioalkyl, thioaryl, substituted thioaryl, thiocycloalkyl,substituted thiocycloalkyl, thioheteroaryl, substituted thioheteroaryl,thioheterocyclic, substituted thioheterocyclic, heteroaryl, substitutedaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic,cycloalkoxy, substituted cycloalkoxy, heteroaryloxy, substitutedheteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy,oxycarbonylamino, oxythiocarbonylamino, cycloalkyloxy, substitutedcycloalkyloxy, heteroaryloxy, substituted heteroaryloxy, —OS(O)₂-alkyl,—OS(O)₂-substituted alkyl, —OS(O)₂-aryl, —OS(O)₂-substituted aryl,—OS(O)₂-heteroaryl, —OS(O)₂-substituted heteroaryl,—OS(O)₂-heterocyclic, —OS(O)₂-substituted heterocyclic, —OSO₂—NRR whereR is hydrogen or alkyl, —NRS(O)₂-alkyl, —NRS(O)₂-substituted alkyl,—NRS(O)₂-aryl, —NRS(O)₂-substituted aryl, —NRS(O)₂-heteroaryl,—NRS(O)₂-substituted heteroaryl, —NRS(O)₂-heterocyclic,—NRS(O)₂-substituted heterocyclic, —NRS(O)₂—NR-alkyl,—NRS(O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl, —NRS(O)₂—NR-substitutedaryl, —NRS(O)₂—NR-heteroaryl, —NRS(O)₂—NR-substituted heteroaryl,—NRS(O)₂—NR-heterocyclic, —NRS(O)₂—NR-substituted heterocyclic where Ris hydrogen or alkyl, mono- and di-alkylamino, mono- and di-(substitutedalkyl)amino, mono and di-arylamino, mono- and di-substituted arylamino,mono- and di-heteroarylamino, mono- and di-substituted heteroarylamino,mono- and di-heterocyclic amino, mono- and di-substituted heterocyclicamino, unsymmetric di-substituted amines having different substituentsindependently selected from the group consisting of alkyl, substitutedalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic and substituted heterocyclic and substituted alkyl groupshaving amino groups blocked by conventional blocking groups such as Boc,Cbz, formyl, and the like or alkyl/substituted alkyl groups substitutedwith —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-alkenyl, —SO₂-substitutedalkenyl, —SO₂-cycloalkyl, —SO₂-substituted cycloalkyl, —SO₂-aryl,—SO₂-substituted aryl, —SO₂-heteroaryl, —SO₂-substituted heteroaryl,—SO₂-heterocyclic, —SO₂-substituted heterocyclic and —SO₂NRR where R ishydrogen or alkyl.

Preferably, the substituents are independently selected from the groupconsisting of alkoxy, substituted alkoxy, acyl, acylamino, acyloxy,amino, substituted amino, aminoacyl, aminocarbonylamino,aminocarbonyloxy, aryl, substituted aryl, aryloxy, substituted aryloxy,carboxyl, carboxyl esters, cyano, cycloalkyl, substituted cycloalkyl,cycloalkyloxy, substituted cycloalkyloxy, halogen, heteroaryl,substituted heteroaryl, heteroaryloxy, substituted heteroaryloxy,heterocyclic, substituted heterocyclic, hydroxyl, nitro, andoxycarbonylamino.

“Alkylene” refers to linear and branched divalent alkyl groups havingfrom 1 to 10 carbon atoms and more preferably 1 to 6 carbon atoms. Thisterm is exemplified by groups such as methylene (—CH₂—), 1,6-heptylene,1,8-octylene, ethylene (—CH₂CH₂—), the propylene isomers (e.g.,—CH₂CH₂CH₂— and —CH(CH₃)CH₂—) and the like.

“Lower alkylene” refers to divalent alkylene groups of from 1 to 4carbon atoms including straight and branched chain alkylene groups. Thisterm is exemplified by groups such as methylene, ethylene, n-propylene,iso-propylene (—CH₂CH(CH₃)— and —CH(CH₃)CH₂—) and the like.

“Substituted alkylene” refers to alkylene groups having from 1 to 5substituents independently selected from the group consisting of alkoxy,substituted alkoxy, acyl, acylamino, thiocarbonylamino, acyloxy, amino,amidino, alkylamidino, thioamidino, aminoacyl, aminocarbonylamino,aminothiocarbonylamino, aminocarbonyloxy, aryl, substituted aryl,aryloxy, substituted aryloxy, aryloxyaryl, substituted aryloxyaryl,halogen, hydroxyl, cyano, nitro, carboxyl, carboxylalkyl,carboxyl-substituted alkyl, carboxyl-cycloalkyl, carboxyl-substitutedcycloalkyl, carboxylaryl, carboxyl-substituted aryl, carboxylheteroaryl,carboxyl-substituted heteroaryl, carboxylheterocyclic,carboxyl-substituted heterocyclic, cycloalkyl, substituted cycloalkyl,guanidino, guanidinosulfone, thiol, thioalkyl, substituted thioalkyl,thioaryl, substituted thioaryl, thiocycloalkyl, substitutedthiocycloalkyl, thioheteroaryl, substituted thioheteroaryl,thioheterocyclic, substituted thioheterocyclic, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, cycloalkoxy,substituted cycloalkoxy, heteroaryloxy, substituted heteroaryloxy,heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino,oxythiocarbonylamino, —OS(O)₂-alkyl, —OS(O)₂-substituted alkyl,—OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,—OS(O)₂-substituted heteroaryl, —OS(O)₂-heterocyclic,—OS(O)₂-substituted heterocyclic, —OSO₂—NRR where R is hydrogen oralkyl, —NRS(O)₂-alkyl, —NRS(O)₂-substituted alkyl, —NRS(O)₂-aryl,—NRS(O)₂-substituted aryl, —NRS(O)₂-heteroaryl, —NRS(O)₂-substitutedheteroaryl, —NRS(O)₂-heterocyclic, —NRS(O)₂-substituted heterocyclic,—NRS(O)₂—NR-alkyl, —NRS(O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl,—NRS(O)₂—NR-substituted aryl, —NRS(O)₂—NR-heteroaryl,—NRS(O)₂—NR-substituted heteroaryl, —NRS(O)₂—NR-heterocyclic,—NRS(O)₂—NR-substituted heterocyclic where R is hydrogen or alkyl, mono-and di-alkylamino, mono- and di-(substituted alkyl)amino, mono- anddi-arylamino, mono- and di-substituted arylamino, mono- anddi-heteroarylamino, mono- and di-substituted heteroarylamino, mono- anddi-heterocyclic amino, mono- and di-substituted heterocyclic amino,unsymmetric di-substituted amines having different substituentsindependently selected from the group consisting of alkyl, substitutedalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic, substituted heterocyclic and substituted alkenyl groupshaving amino groups blocked by conventional blocking groups such as Boc,Cbz, formyl, and the like or alkenyl/substituted alkenyl groupssubstituted with —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-alkenyl,—SO₂-substituted alkenyl, —SO₂-cycloalkyl, —SO₂-substituted cycloalkyl,—SO₂-aryl, —SO₂-substituted aryl, —SO₂-heteroaryl, —SO₂-substitutedheteroaryl, —SO₂-heterocyclic, —SO₂-substituted heterocyclic and —SO₂NRRwhere R is hydrogen or alkyl.

“Alkynyl” refers to alkynyl group preferably having from 2 to 10 carbonatoms and more preferably 3 to 6 carbon atoms and having at least 1 andpreferably from 1-2 sites of alkynyl unsaturation.

“Lower alkynyl” refers to an alkynyl group preferably having from 2 to 6carbon atoms and having at least 1 site and preferably only 1 site ofalkynyl unsaturation (i.e., —C≡C—). This term is exemplified by groupssuch as acetyl (—C≡CH), propargyl (—CH₂—C≡CH), 3-butynyl (—CH₂CH₂C≡CH₃)and the like.

“Substituted alkynyl” refers to alkynyl groups having from 1 to 5substituents independently selected from the group consisting of alkoxy,substituted alkoxy, acyl, acylamino, thiocarbonylamino, acyloxy, amino,amidino, alkylamidino, thioamidino, aminoacyl, aminocarbonylamino,aminothiocarbonylamino, aminocarbonyloxy, aryl, substituted aryl,aryloxy, substituted aryloxy, aryloxyaryl, substituted aryloxyaryl,halogen, hydroxyl, cyano, nitro, carboxyl, carboxylalkyl,carboxyl-substituted alkyl, carboxylcycloalkyl, carboxyl-substitutedcycloalkyl, carboxylaryl, carboxyl-substituted aryl, carboxylheteroaryl,carboxyl-substituted heteroaryl, carboxylheterocyclic,carboxyl-substituted heterocyclic, cycloalkyl, substituted cycloalkyl,guanidino, guanidinosulfone, thiol, thioalkyl, substituted thioalkyl,thioaryl, substituted thioaryl, thiocycloalkyl, substitutedthiocycloalkyl, thioheteroaryl, substituted thioheteroaryl,thioheterocyclic, substituted thioheterocyclic, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, cycloalkoxy,substituted cycloalkoxy, heteroaryloxy, substituted heteroaryloxy,heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino,oxythiocarbonylamino, —OS(O)₂-alkyl, —OS(O)₂-substituted alkyl,—OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,—OS(O)₂-substituted heteroaryl, —OS(O)₂-heterocyclic,—OS(O)₂-substituted heterocyclic, —OSO₂—NRR where R is hydrogen oralkyl, —NRS(O)₂-alkyl, —NRS(O)₂-substituted alkyl, —NRS(O)₂-aryl,—NRS(O)₂-substituted aryl, —NRS(O)₂-heteroaryl, —NRS(O)₂-substitutedheteroaryl, —NRS(O)₂-heterocyclic, —NRS(O)₂-substituted heterocyclic,—NRS(O)₂—NR-alkyl, —NRS(O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl,—NRS(O)₂—NR-substituted aryl, —NRS(O)₂—NR-heteroaryl,—NRS(O)₂—NR-substituted heteroaryl, —NRS(O)₂—NR-heterocyclic,—NRS(O)₂—NR-substituted heterocyclic where R is hydrogen or alkyl, mono-and di-alkylamino, mono- and di-(substituted-alkyl)amino, mono- anddi-arylamino, mono- and di-substituted arylamino, mono- anddi-heteroarylamino, mono- and di-substituted heteroarylamino, mono- anddi-heterocyclic amino, mono- and di-substituted heterocyclic amino,unsymmetric di-substituted amines having different substituentsindependently selected from the group consisting of alkyl, substitutedalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic, substituted heterocyclic and substituted alkynyl groupshaving amino groups blocked by conventional blocking groups such as Boc,Cbz, formyl, and the like or alkynyl/substituted alkynyl groupssubstituted with —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-alkenyl,—SO₂-substituted alkenyl, —SO₂-cycloalkyl, —SO₂-substituted cycloalkyl,—SO₂-aryl, —SO₂-substituted aryl, —SO₂-heteroaryl, —SO₂-substitutedheteroaryl, —SO₂-heterocyclic, —SO₂-substituted heterocyclic and —SO₂NRRwhere R is hydrogen or alkyl.

“Amidino” refers to the group H₂NC(═NH)— and the term “alkylamidino”refers to compounds having 1 to 3 alkyl groups (e.g., alkylHNC(═NH)—).

“Amino” refers to the group —NH₂.

“Substituted amino” refers to the group —NRR, where each R group isindependently selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl, heterocyclic, substitutedheterocyclic, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-alkenyl,—SO₂-substituted alkenyl, —SO₂-cycloalkyl, —SO₂-substituted cycloalkyl,—SO₂-aryl, —SO₂-substituted aryl, —SO₂-heteroaryl, —SO₂-substitutedheteroaryl, —SO₂-heterocyclic, —SO₂-substituted heterocyclic, providedthat both R groups are not hydrogen; or the R groups can be joinedtogether with the nitrogen atom to form a heterocyclic or substitutedheterocyclic ring.

“Aminoacyl” refers to the groups —NRC(O)alkyl, —NRC(O)substituted alkyl,—NRC(O)cycloalkyl, —NRC(O)substituted cycloalkyl, —NRC(O)alkenyl,—NRC(O)substituted alkenyl, —NRC(O)alkynyl, —NRC(O)substituted alkynyl,—NRC(O)aryl, —NRC(O)substituted aryl, —NRC(O)heteroaryl,—NRC(O)substituted heteroaryl, —NRC(O)heterocyclic, and—NRC(O)substituted heterocyclic where R is hydrogen or alkyl and whereinalkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic andsubstituted heterocyclic are as defined herein.

“Aminocarbonylamino” refers to the groups —NRC(O)NRR, —NRC(O)NR-alkyl,—NRC(O)NR-substituted alkyl, —NRC(O)NR-alkenyl, —NRC(O)NR-substitutedalkenyl, —NRC(O)NR-alkynyl, —NRC(O)NR-substituted alkynyl,—NRC(O)NR-aryl, —NRC(O)NR-substituted aryl, —NRC(O)NR-cycloalkyl,—NRC(O)NR-substituted cycloalkyl, —NRC(O)NR-heteroaryl, and—NRC(O)NR-substituted heteroaryl, —NRC(O)NR-heterocyclic, and—NRC(O)NR-substituted heterocyclic where each R is independentlyhydrogen, alkyl or where each R is joined to form together with thenitrogen atom a heterocyclic or substituted heterocyclic ring as well aswhere one of the amino groups is blocked by conventional blocking groupssuch as Boc, Cbz, formyl, and the like and wherein alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic and substituted heterocyclic are asdefined herein.

“Aminocarbonyloxy” refers to the groups —NRC(O)O-alkyl,—NRC(O)O-substituted alkyl, —NRC(O)O-alkenyl, —NRC(O)O-substitutedalkenyl, —NRC(O)O-alkynyl, —NRC(O)O-substituted alkynyl,—NRC(O)O-cycloalkyl, —NRC(O)O-substituted cycloalkyl, —NRC(O)O-aryl,—NRC(O)O-substituted aryl, —NRC(O)O-heteroaryl, —NRC(O)O-substitutedheteroaryl, —NRC(O)O-heterocyclic, and —NRC(O)O-substituted heterocyclicwhere R is hydrogen or alkyl and wherein alk-yl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Aminosulfonyl” refers to the groups —NRSO₂alkyl, —NRSO₂substitutedalkyl, —NRSO₂cycloalkyl, —NRSO₂substituted cycloalkyl, —NRSO₂alkenyl,—NRSO₂substituted alkenyl, —NRSO₂alkynyl, —NRSO₂substituted alkynyl,—NRSO₂aryl, —NRSO₂substituted aryl, —NRSO₂heteroaryl, —NRSO₂substitutedheteroaryl, —NRSO₂heterocyclic, and —NRSO₂substituted heterocyclic whereR is hydrogen or alkyl and wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Aminosulfonylamino” refers to the groups —NRSO₂NRR, —NRSO₂NR-alkyl,—NRSO₂NR-substituted alkyl, —NRSO₂NR-alkenyl, —NRSO₂NR-substitutedalkenyl, —NRSO₂NR-alkynyl, —NRSO₂NR-substituted alkynyl, —NRSO₂NR-aryl,—NRSO₂NR-substituted aryl, —NRSO₂NR-cycloalkyl, —NRSO₂NR-substitutedcycloalkyl, —NRSO₂NR-heteroaryl, and —NRSO₂NR-substituted heteroaryl,—NRSO₂NR-heterocyclic, and —NRSO₂NR-substituted heterocyclic, where eachR is independently hydrogen, alkyl or where each R is joined to formtogether with the nitrogen atom a heterocyclic or substitutedheterocyclic ring as well as where one of the amino groups is blocked byconventional blocking groups such as Boc, Cbz, formyl, and the like andwherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic andsubstituted heterocyclic are as defined herein.

“Aminosulfonyloxy” refers to the groups —NRSO₂O-alkyl,—NRSO₂O-substituted alkyl, —NRSO₂O-alkenyl, —NRSO₂O-substituted alkenyl,—NRSO₂O-alkynyl, —NRSO₂O-substituted alkynyl, —NRSO₂O-cycloalkyl,—NRSO₂O-substituted cycloalkyl, —NRSO₂O-aryl, —NRSO₂O-substituted aryl,—NRSO₂O-heteroaryl, —NRSO₂O-substituted heteroaryl,—NRSO₂O-heterocyclic, and —NRSO₂O-substituted heterocyclic where R ishydrogen or alkyl and wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Aminothiocarbonylamino” refers to the groups —NRC(S)NRR,—NRC(S)NR-alkyl, —NRC(S)NR-substituted alkyl, —NRC(S)NR-alkenyl,—NRC(S)NR-substituted alkenyl, —NRC(S)NR-alkynyl, —NRC(S)NR-substitutedalkynyl, —NRC(S)NR-aryl, —NRC(S)NR-substituted aryl,—NRC(S)NR-cycloalkyl, —NRC(S)NR-substituted cycloalkyl,—NRC(S)NR-heteroaryl, and —NRC(S)NR-substituted heteroaryl,—NRC(S)NR-heterocyclic, and —NRC(S)NR-substituted heterocyclic whereeach R is independently hydrogen, alkyl or where each R is joined toform together with the nitrogen atom a heterocyclic or substitutedheterocyclic ring as well as where one of the amino groups is blocked byconventional blocking groups such as Boc, Cbz, formyl, and the like andwherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic andsubstituted heterocyclic are as defined herein.

“Aryl” or “Ar” refers to an unsaturated aromatic carbocyclic group offrom 6 to 14 carbon atoms having a single ring (e.g., phenyl) ormultiple condensed rings (e.g., naphthyl or anthryl) which condensedrings may or may not be aromatic (e.g., 2-benzoxazolinone,2H-1,4-benzoxazin-3(4H)-one-7yl, and the like) provided that the pointof attachment is through an aromatic ring atom. Preferred aryls includephenyl, naphthyl and 5,6,7,8-tetrahydronaphth-2-yl.

“Substituted aryl” refers to aryl groups which are substituted with from1 to 3 substituents selected from the group consisting of hydroxy, acyl,acylamino, thiocarbonylamino, acyloxy, alkyl, substituted alkyl, alkoxy,substituted alkoxy, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, amidino, alkylamidino, thioamidino, amino, aminoacyl,aminocarbonyloxy, aminocarbonylamino, aminothiocarbonylamino, aryl,substituted aryl, aryloxy, substituted aryloxy, cycloalkoxy, substitutedcycloalkoxy, heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy,substituted heterocyclyloxy, carboxyl, carboxylalkyl,carboxyl-substituted alkyl, carboxyl-cycloalkyl, carboxyl-substitutedcycloalkyl, carboxylaryl, carboxyl-substituted aryl, carboxylheteroaryl,carboxyl-substituted heteroaryl, carboxylheterocyclic,carboxyl-substituted heterocyclic, carboxylamido, cyano, thiol,thioalkyl, substituted thioalkyl, thioaryl, substituted thioaryl,thioheteroaryl, substituted thioheteroaryl, thiocycloalkyl, substitutedthiocycloalkyl, thioheterocyclic, substituted thioheterocyclic,cycloalkyl, substituted cycloalkyl, guanidino, guanidinosulfone, halo,nitro, heteroaryl, substituted heteroaryl, heterocyclic, substitutedheterocyclic, cycloalkoxy, substituted cycloalkoxy, heteroaryloxy,substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy,oxycarbonylamino, oxythiocarbonylamino, —S(O)₂-alkyl, —S(O)₂-substitutedalkyl, —S(O)₂-cycloalkyl, —S(O)₂-substituted cycloalkyl, —S(O)₂-alkenyl,—S(O)₂-substituted alkenyl, —-S(O)₂-aryl, —S(O)₂-substituted aryl,—S(O)₂-heteroaryl, —S(O)₂-substituted heteroaryl, —S(O)₂-heterocyclic,—S(O)₂-substituted heterocyclic, —OS(O)₂-alkyl, —OS(O)₂-substitutedalkyl, —OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,—OS(O)2-substituted heteroaryl, —OS(O)₂-heterocyclic,—OS(O)₂-substituted heterocyclic, —OSO₂—NRR where R is hydrogen oralkyl, —NRS (O)₂-alkyl, —NRS (O)₂-substituted alkyl, —NRS (O)₂-aryl,—NRS(O)₂-substituted aryl, —NRS (O)₂-heteroaryl, —NRS (O)₂-substitutedheteroaryl, —NRS(O)₂-heterocyclic, —NRS (O)₂-substituted heterocyclic,—NRS (O)₂—NR-alkyl, —NRS (O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl,—NRS (O)₂—NR-substituted aryl, —NRS (O)₂—NR-heteroaryl,—NRS(O)₂—NR-substituted heteroaryl, —NRS(O)₂—NR-heterocyclic, —NRS(O)₂—NR-substituted heterocyclic where R is hydrogen or alkyl, mono- anddi-alkylamino, mono- and di-(substituted alkyl)amino, mono- anddi-arylamino, mono- and di-substituted arylamino, mono- anddi-heteroarylamino, mono- and di-substituted heteroarylamino, mono- anddi-heterocyclic amino, mono- and di-substituted heterocyclic amino,unsymmetric di-substituted amines having different substituentsindependently selected from the group consisting of alkyl, substitutedalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic and substituted heterocyclic and amino groups on thesubstituted aryl blocked by conventional blocking groups such as Boc,Cbz, formyl, and the like or substituted with —SO₂NRR where R ishydrogen or alkyl.

Preferred substituents are selected from the group consisting ofhydroxy, acyl, acylamino, acyloxy, alkyl, substituted alkyl, alkoxy,substituted alkoxy, alkenyl, substituted alkenyl, amino, substitutedamino, aminoacyl, aminocarbonyloxy, aminocarbonylamino, aryl,substituted aryl, aryloxy, substituted aryloxy, cycloalkoxy, substitutedcycloalkoxy, heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy,substituted heterocyclyloxy, carboxyl, carboxyl esters, cyano,cycloalkyl, substituted cycloalkyl, halo, nitro, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, andoxycarbonylamino.

“Aryloxy” refers to the group aryl-O— which includes, by way of example,phenoxy, naphthoxy, and the like.

“Substituted aryloxy” refers to substituted aryl-O— groups.

“Aryloxyaryl” refers to the group -aryl-O-aryl.

“Substituted aryloxyaryl” refers to aryloxyaryl groups substituted withfrom 1 to 3 substituents on either or both aryl rings selected from thegroup consisting of hydroxy, acyl, acylamino, thiocarbonylamino,acyloxy, alkyl, substituted alkyl, alkoxy, substituted alkoxy, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, amidino,alkylamidino, thioamidino, amino, aminoacyl, aminocarbonyloxy,aminocarbonylamino, aminothiocarbonylamino, aryl, substituted aryl,aryloxy, substituted aryloxy, cycloalkoxy, substituted cycloalkoxy,heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy, substitutedheterocyclyloxy, carboxyl, carboxylalkyl, carboxyl-substituted alkyl,carboxyl-cycloalkyl, carboxyl-substituted cycloalkyl, carboxylaryl,carboxyl-substituted aryl, carboxylheteroaryl, carboxyl-substitutedheteroaryl, carboxylheterocyclic, carboxyl-substituted heterocyclic,carboxylamido, cyano, thiol, thioalkyl, substituted thioalkyl, thioaryl,substituted thioaryl, thioheteroaryl, substituted thioheteroaryl,thiocycloalkyl, substituted thiocycloalkyl, thioheterocyclic,substituted thioheterocyclic, cycloalkyl, substituted cycloalkyl,guanidino, guanidinosulfone, halo, nitro, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, cycloalkoxy,substituted cycloalkoxy, heteroaryloxy, substituted heteroaryloxy,heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino,oxythiocarbonylamino, —S(O)₂-alkyl, —S(O)₂-substituted alkyl,—S(O)2-cycloalkyl, —S(O)₂-substituted cycloalkyl, —S(O)₂-alkenyl,—S(O)₂-substituted alkenyl, —S(O)₂-aryl, —S(O)₂-substituted aryl,—S(O)₂-heteroaryl, —S(O)₂-substituted heteroaryl, —S(O)₂-heterocyclic,—S(O)₂-substituted heterocyclic, —OS(O)₂-alkyl, —OS(O)₂-substitutedalkyl, —OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,—OS(O)₂-substituted heteroaryl, —OS(O)₂-heterocyclic,—OS(O)₂-substituted heterocyclic, —OSO₂—NRR where R is hydrogen oralkyl, —NRS(O)₂-alkyl, —NRS(O)₂-substituted alkyl, —NRS(O)₂-aryl,—NRS(O)₂-substituted aryl, —NRS(O)₂-heteroaryl, —NRS(O)₂-substitutedheteroaryl, —NRS(O)₂-heterocyclic, —NRS(O)₂-substituted heterocyclic,—NRS(O)₂—NR-alkyl, —NRS(O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl,—NRS(O)₂—NR-substituted aryl, —NRS(O)₂—NR-heteroaryl,—NRS(O)₂—NR-substituted heteroaryl, —NRS(O)₂—NR-heterocyclic,—NRS(O)₂—NR-substituted heterocyclic where R is hydrogen or alkyl, mono-and di-alkylamino, mono- and di-(substituted alkyl)amino, mono- anddi-arylamino, mono- and di-substituted arylamino, mono- anddi-heteroarylamino, mono- and di-substituted heteroarylamino, mono- anddi-heterocyclic amino, mono- and di-substituted heterocyclic amino,unsymmetric di-substituted amines having different substituentsindependently selected from the group consisting of alkyl, substitutedalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic and substituted heterocyclic and amino groups on thesubstituted aryl blocked by conventional blocking groups such as Boc,Cbz, formyl, and the like or substituted with —SO₂NRR where R ishydrogen or alkyl.

“Aralkoxy” refers to aryl-alkylene-O— groups.

“Substituted aralkoxy” refers to substituted aryl-alkylene-O— groups.

“Carboxyl” refers to the group —COOH and pharmaceutically acceptablesalts thereof.

“Carboxyl esters” refers —C(O)O-alkyl, —C(O)O-substituted alkyl,—C(O)O-alkenyl, —C(O)O-substituted alkenyl, —C(O)O-aryl,—C(O)O-substituted aryl, —C(O)O-cycloalkyl, —C(O)O-substitutedcycloalkyl, —C(O)O-heteroaryl, —C(O)O-substituted heteroaryl,—C(O)O-heterocyclic, and —C(O)O-substituted heterocyclic.

“Cycloalkenyl” refers to cyclic alkenyl groups of from 3 to 8 carbonatoms having single or multiple unsaturation but which are not aromatic.

“Cycloalkoxy” refers to —O-cycloalkylgroups.

“Substituted cycloalkoxy” refers to —O-substituted cycloalkyl groups.

“Cycloalkyl”, with regard to the compounds of Formulae I and II and thePEG derivatives, refers to cyclic alkyl groups of from 3 to 12 carbonatoms having a single or multiple condensed rings including, by way ofexample, adamantyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cyclooctyl and the like. Preferably “cycloalkyl” refers to cyclic alkylgroups of from 3 to 8 carbon atoms having a single cyclic ring.

“Cycloalkyl”, with regards to the compounds of Formulae III-IX, refersto cyclic alkyl groups of from 3 to 8 carbon atoms having a singlecyclic ring including, by way of example,cyclopropyl-cyclobutyl-cyclopentyl,-cyclohexyl, cyclooctyl and the like.Excluded from this definition are multi-ring alkyl groups such asadamantanyl, etc.

“Lower cycloalkyl” refers to cyclic alkyl groups of from 3 to 6 carbonatoms having a single cyclic ring including, by way of example,cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

“Substituted cycloalkyl” and “substituted cycloalkenyl” refers to acycloalkyl or cycloalkenyl group, preferably of from 3 to 8 carbonatoms, having from 1 to 5 substituents independently selected from thegroup consisting of oxo (═O), thioxo (═S), alkoxy, substituted alkoxy,acyl, acylamino, thiocarbonylamino, acyloxy, amino, amidino,alkylamidino, thioamidino, aminoacyl, aminocarbonylamino,aminothiocarbonylamino, aminocarbonyloxy, aryl, substituted aryl,aryloxy, substituted aryloxy, aryloxyaryl, substituted aryloxyaryl,halogen, hydroxyl, cyano, nitro, carboxyl, carboxylalkyl,carboxyl-substituted alkyl, carboxyl-cycloalkyl, carboxyl-substitutedcycloalkyl, carboxylaryl, carboxyl-substituted aryl, carboxylheteroaryl,carboxyl-substituted heteroaryl, carboxylheterocyclic,carboxyl-substituted heterocyclic, cycloalkyl, substituted cycloalkyl,guanidino, guanidinosulfone, thiol, thioalkyl, substituted thioalkyl,thioaryl, substituted thioaryl, thiocycloalkyl, substitutedthiocycloalkyl, thioheteroaryl, substituted thioheteroaryl,thioheterocyclic, substituted thioheterocyclic, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, cycloalkoxy,substituted cycloalkoxy, heteroaryloxy, substituted heteroaryloxy,heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino,oxythiocarbonylamino, —OS(O)₂-alkyl, —OS(O)₂-substituted alkyl,—OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,—OS(O)₂-substituted heteroaryl, —OS(O)₂-heterocyclic,—OS(O)₂-substituted heterocyclic, —OSO₂—NRR where R is hydrogen oralkyl, —NRS(O)₂-alkyl, —NRS(O)₂-substituted alkyl, —NRS(O)₂-aryl,—NRS(O)₂-substituted aryl, —NRS(O)₂-heteroaryl, —NRS(O)₂-substitutedheteroaryl, —NRS(O)₂-heierocyclic, —NRS(O)₂-substituted heterocyclic,—NRS(O)₂—NR-alkyl, —NRS(O)₂-NR-substituted alkyl, —NRS(O)₂—NR-aryl,—NRS(O)₂—NR-substituted aryl, —NRS(O)₂—NR-heteroaryl,—NRS(O)₂—NR-substituted heteroaryl, —NRS(O)₂—NR-heterocyclic,—NRS(O)₂—NR-substituted heterocyclic where R is hydrogen or alkyl, mono-and di-alkylamino, mono- and di-(substituted alkyl)amino, mono- anddi-arylamino, mono- and di-substituted arylamino, mono- anddi-heteroarylamino, mono- and di-substituted heteroarylamino, mono- anddi-heterocyclic amino, mono- and di-substituted-heterocyclic amino,unsymmetric di-substituted amines having different substituentsindependently selected from the group consisting of alkyl, substitutedalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic and substituted heterocyclic and substituted alkynyl groupshaving amino groups blocked by conventional blocking groups such as Boc,Cbz, formyl, and the like or alkynyl/substituted alkynyl groupssubstituted with —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-alkenyl,—SO₂-substituted alkenyl, —SO₂-cycloalkyl, —SO₂-substituted cycloalkyl,—SO₂-aryl, —SO₂-substituted aryl, —SO₂-heteroaryl, —SO₂-substitutedheteroaryl, —SO₂-heterocyclic, —SO₂-substituted heterocyclic and —SO₂NRRwhere R is hydrogen or alkyl.

Preferred substituents are selected from the group consisting of oxo(═O), thioxo (═S), alkoxy, substituted alkoxy, acyl, acylamino, acyloxy,amino, substituted amino, aminoacyl, aminocarbonylamino,aminocarbonyloxy, aryl, substituted aryl, aryloxy, substituted aryloxy,carboxyl, carboxyl esters, cyano, cycloalkyl, substituted cycloalkyl,cycloalkyloxy, substituted cycloalkyloxy, halogen, heteroaryl,substituted heteroaryl, heteroaryloxy, substituted heteroaryloxy,heterocyclic, substituted heterocyclic, hydroxyl, nitro, andoxycarbonylamino.

“Guanidino” refers to the groups —NRC(═NR)NRR, —NRC(═NR)NR-alkyl,—NRC(═NR)NR-substituted alkyl, —NRC(═NR)NR-alkenyl,—NRC(═NR)NR-substituted alkenyl, —NRC(═NR)NR-alkynyl,—NRC(═NR)NR-substituted alkynyl, —NRC(═NR)NR-aryl,—NRC(═NR)NR-substituted aryl, —NRC(═NR)NR-cycloalkyl,—NRC(═NR)NR-heteroaryl, —NRC(═NR)NR-substituted heteroaryl,—NRC(═NR)NR-heterocyclic, and —NRC(═NR)NR-substituted heterocyclic whereeach R is independently hydrogen and alkyl as well as where one of theamino groups is blocked by conventional blocking groups such as Boc,Cbz, formyl, and the like and wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Guanidinosulfone” refers to the groups —NRC(═NR)NRSO₂-alkyl,—NRC(═NR)NRSO₂-substituted alkyl, —NRC(═NR)NRSO₂-alkenyl,—NRC(═NR)NRSO₂-substituted alkenyl, —NRC(═NR)NRSO₂-alkynyl,—NRC(═NR)NRSO₂-substituted alkynyl, —NRC(═NR)NRSO₂-aryl,—NRC(═NR)NRSO₂-substituted aryl,-—NRC(═NR)NRSO₂cycloalkyl,—NRC(═NR)NRSO₂-substituted cycloalkyl, —NRC(═NR)NRSO₂-heteroaryl, and—NRC(═NR)NRSO₂-substituted heteroaryl, —NRC(═NR)NRSO₂-heterocyclic, and—NRC(═NR)NRSO₂-substituted heterocyclic where each R is independentlyhydrogen and alkyl and wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Halo” or “halogen” refers to fluoro, chloro, bromo and iodo andpreferably is fluoro, chloro or bromo.

“Heteroaryl” refers to an aromatic carbocyclic group of from 2 to 10carbon atoms and 1 to 4 heteroatoms selected from the group consistingof oxygen, nitrogen and sulfur within the ring or oxides thereof. Suchheteroaryl groups can have a single ring (e.g., pyridyl or furyl) ormultiple condensed rings (e.g., indolizinyl or benzothienyl) wherein oneor more of the condensed rings may or may not be aromatic provided thatthe point of attachment is through an aromatic ring atom. Additionally,the heteroatoms of the heteroaryl group may be oxidized, i.e., to formpyridine N-oxides or 1,1-dioxo-1,2,5-thiadiazoles and the like.Additionally, the carbon atoms of the ring may be substituted with anoxo (═O). Preferred heteroaryls include pyridyl, pyrrolyl, indolyl,furyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1-oxo-1,2,5-thiadiazolyl and1,1-dioxo-1,2,5-thiadiazolyl. The term “heteroaryl having two nitrogenatoms in the heteroaryl ring” refers to a heteroaryl group having two,and only two, nitrogen atoms in the heteroaryl ring and optionallycontaining 1 or 2 other heteroatoms in the heteroaryl ring, such asoxygen or sulfur.

“Substituted heteroaryl” refers to heteroaryl groups which aresubstituted with from 1 to 3 substituents selected from the groupconsisting of hydroxy, acyl, acylamino, thiocarbonylamino, acyloxy,alkyl, substituted alkyl, alkoxy, substituted alkoxy, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, amidino,alkylamidino, thioamidino, amino, aminoacyl, aminocarbonyloxy,aminocarbonylamino, aminothiocarbonylamino, aryl, substituted aryl,aryloxy, substituted aryloxy, cycloalkoxy, substituted cycloalkoxy,heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy, substitutedheterocyclyloxy, carboxyl, carboxylalkyl, carboxyl-substituted alkyl,carboxyl-cycloalkyl, carboxyl-substituted cycloalkyl, carboxylaryl,carboxyl-substituted aryl, carboxylheteroaryl, carboxyl-substitutedheteroaryl, carboxylheterocyclic, carboxyl-substituted heterocyclic,carboxylamido, cyano, thiol, thioalkyl, substituted thioalkyl, thioaryl,substituted thioaryl, thioheteroaryl, substituted thioheteroaryl,thiocycloalkyl, substituted thiocycloalkyl, thioheterocyclic,substituted thioheterocyclic, cycloalkyl, substituted cycloalkyl,guanidino, guanidinosulfone, halo, nitro, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, cycloalkoxy,substituted cycloalkoxy, heteroaryloxy, substituted heteroaryloxy,heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino,oxythiocarbonylamino, —S(O)₂-alkyl, —S(O)2-substituted alkyl,—S(O)₂-cycloalkyl, —S(O)₂-substituted cycloalkyl, —S(O)₂-alkenyl,—S(O)₂-substituted alkenyl, —S(O)₂-aryl, —S(O)₂-substituted aryl,—S(O)₂-heteroaryl, —S(O)₂-substituted heteroaryl, —S(O)₂-heterocyclic,—S(O)₂-substituted heterocyclic, —OS(O)₂-alkyl, —OS(O)₂-substitutedalkyl, —OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,—OS(O)₂-substituted heteroaryl, —OS(O)₂-heterocyclic,—OS(O)₂-substituted heterocyclic, —OSO₂—NRR where R is hydrogen oralkyl, —NRS(O)₂-alkyl, —NRS(O)₂-substituted alkyl, —NRS(O)₂-aryl,—NRS(O)₂-substituted aryl, —NRS (O)₂-heteroaryl, —NRS(O)₂-substitutedheteroaryl, —NRS (O)₂-heterocyclic, —NRS(O)₂-substituted heterocyclic,—NRS(O)₂—NR-alkyl, —NRS(O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl,—NRS(O)₂—NR-substituted aryl, —NRS (O)₂—NR-heteroaryl,—NRS(O)₂—NR-substituted heteroaryl, —NRS(O)₂—NR-heterocyclic,—NRS(O)₂—NR-substituted heterocyclic where R is hydrogen or alkyl, mono-and di-alkylamino, mono- and di-(substituted alkyl)amino, mono- anddi-arylamino, mono- and di-substituted arylamino, mono- anddi-heteroarylamino, mono- and di-substituted heteroarylamino, mono- anddi-heterocyclic amino, mono- and di-substituted heterocyclic amino,unsymmetric di-substituted amines having different substituentsindependently selected from the group consisting of alkyl, substitutedalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic and substituted heterocyclic and amino groups on thesubstituted aryl blocked by conventional blocking groups such as Boc,Cbz, formyl, and the like or substituted with —SO₂NRR where R ishydrogen or alkyl.

Preferably the substituents are selected from the group consisting ofthose defined above as preferred for substituted aryl.

“Heteroaryloxy” refers to the group —O-heteroaryl and “substitutedheteroaryloxy” refers to the group —O-substituted heteroaryl.

“Heteroaralkoxy” refers to the group heteroaryl-alkylene-O—.

“Substituted heteroaralkoxy” refers to the group substitutedheteroaryl-alkylene-O—.

“Heterocycle” or “heterocyclic” refers to a saturated or unsaturatedgroup having a single ring or multiple condensed rings, from 1 to 10carbon atoms and from 1 to 4 hetero atoms selected from the groupconsisting of nitrogen, sulfur or oxygen within the ring wherein, infused ring systems, one or more the rings can be aryl or heteroaryl.

“Substituted heterocyclic” refers to heterocycle groups which aresubstituted with from 1 to 3 substituents selected from the groupconsisting of oxo (═O), thioxo (═S), alkoxy, substituted alkoxy, acyl,acylamino, thiocarbonylamino, acyloxy, amino, amidino, alkylamidino,thioamidino, aminoacyl, aminocarbonylamino, aminothiocarbonylamino,aminocarbonyloxy, aryl, substituted aryl, aryloxy, substituted aryloxy,aryloxyaryl, substituted aryloxyaryl, halogen, hydroxyl, cyano, nitro,carboxyl, carboxylalkyl, carboxyl-substituted alkyl,carboxyl-cycloalkyl, carboxyl-substituted cycloalkyl, carboxylaryl,carboxyl-substituted aryl, carboxylheteroaryl, carboxyl-substitutedheteroaryl, carboxylheterocyclic, carboxyl-substituted heterocyclic,cycloalkyl, substituted cycloalkyl, guanidino, guanidinosulfone, thiol,thioalkyl, substituted thioalkyl, thioaryl, substituted thioaryl,thiocycloalkyl, substituted thiocycloalkyl, thioheteroaryl, substitutedthioheteroaryl, thioheterocyclic, substituted thioheterocyclic,heteroaryl, substituted heteroaryl, heterocyclic, substitutedheterocyclic, cycloalkoxy, substituted cycloalkoxy, heteroaryloxy,substituted heteroaryloxy, —C(O)O-aryl, —C(O)O-substituted aryl,heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino,oxythiocarbonylamino, —OS(O)₂-alkyl, —OS(O)₂-substituted alkyl,—OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,—OS(O)₂-substituted heteroaryl, —OS(O)₂-heterocyclic,—OS(O)₂-substituted heterocyclic, —OSO₂—NRR where R is hydrogen oralkyl, —NRS(O)₂-alkyl, —NRS(O)₂-substituted alkyl, —NRS(O)₂-aryl,—NRS(O)₂-substituted aryl, —NRS(O)₂-heteroaryl, —NRS(O)₂-substitutedheteroaryl, —NRS(O)₂-heterocyclic, —NRS(O)₂-substituted heterocyclic,—NRS(O)₂—NR-alkyl, —NRS(O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl,—NRS(O)₂—NR-substituted aryl, —NRS(O)₂—NR-heteroaryl,—NRS(O)₂—NR-substituted-heteroaryl, —NRS(O)₂—NR-heterocyclic,—NRS(O)₂—NR-substituted heterocyclic where R is hydrogen or alkyl, mono-and di-alkylamino, mono- and di-(substituted alkyl)amino, mono- anddi-arylamino, mono- and di-substituted arylamino, mono- anddi-heteroarylamino, mono- and di-substituted heteroarylamino, mono- anddi-heterocyclic amino, mono- and di-substituted heterocyclic amino,unsymmetric di-substituted amines having different substituentsindependently selected from the group consisting of alkyl, substitutedalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic and substituted heterocyclic and substituted alkynyl groupshaving amino groups blocked by conventional blocking groups such as Boc,Cbz, formyl, and the like or alkynyl/substituted alkynyl groupssubstituted with —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-alkenyl,—SO₂-substituted alkenyl, —SO₂-cycloalkyl, —SO₂-substituted cycloalkyl,—SO2-aryl, —SO₂-substituted aryl, —SO₂-heteroaryl, —SO₂-substitutedheteroaryl, —SO₂-heterocyclic, —SO₂-substituted heterocyclic and —SO₂NRRwhere R is hydrogen or alkyl.

Preferably, the substituents are selected from the group consisting ofthe preferred substitutents defined for substituted cycloalkyl.

Examples of heterocycles and heteroaryls include, but are not limitedto, azetidine, pyrrole, imidazole, pyrazole, pyridine, pyrazine,pyrimidine, pyridazine, indolizine, isoindole, indole, dihydroindole,indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine,naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine,carbazole, carboline, phenanthridine, acridine, phenanthroline,isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine,imidazolidine, imidazoline, piperidine, piperazine, indoline,phthalimide, 1,2,3,4-tetrahydroisoquinoline,4,5,6,7-tetrahydrobenzo[b]thiophene, thiazole, thiazolidine, thiophene,benzo[b]thiophene, morpholino, morpholinyl, thiomorpholino,thiomorpholinyl (also referred to as thiamorpholinyl), piperidinyl,pyrrolidine, tetrahydrofuranyl, and the like.

“Heterocyclyloxy” refers to the group -0-heterocyclic and “substitutedheterocyclyloxy” refers to the group —O-substituted heterocyclic.

“Lower alkylenecycloalkyl” refers to the group consisting of a loweralkylene-lower cycloalkyl, as defined herein. Such groups areexemplified by methylenecyclopropyl (—CH₂-cyclopropyl),ethylenecyclopropyl and the like.

“N,N-Dimethylcarbamyloxy” refers to the group —OC(O)N(CH₃)₂.

“Oxo” refers to (═O).

“Oxyalkylene” refers to —OCH₂CHR^(d)— where R^(d) is alkyl.

“Oxycarbonylamino” refers to the groups —OC(O)NH₂, —OC(O)NRR,—OC(O)NR-alkyl, —OC(O)NR-substituted alkyl, —OC(O)NR-alkenyl,—OC(O)NR-substituted alkenyl, —OC(O)NR-alkynyl, —OC(O)NR-substitutedalkynyl, —OC(O)NR-cycloalkyl, —OC(O)NR-substituted cycloalkyl,—OC(O)NR-aryl, —OC(O)NR-substituted aryl, —OC(O)NR-heteroaryl,—OC(O)NR-substituted heteroaryl, —OC(O)NR-heterocyclic, and—OC(O)NR-substituted heterocyclic where R is hydrogen, alkyl or whereeach R is joined to form, together with the nitrogen atom a heterocyclicor substituted heterocyclic ring and wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Oxysulfonyl” refers to the groups alkyl—SO₂O—, substituted alkyl—SO₂O—,alkenyl-SO₂O—, substituted alkenyl—SO₂O—, alkynyl—SO₂O—, substitutedalkynyl—SO₂O—, aryl—SO₂O—, substituted aryl—SO₂O—, cycloalkyl—SO₂O—,substituted cycloalkyl—SO₂O—, heteroaryl—SO₂O—, substitutedheteroaryl—SO₂O—, heterocyclic—SO₂O—, and substitutedheterocyclic—SO₂O—wherein alkyl, substituted alkyl, alkenyl, substitutedalkenyl, alkynyl, substituted alkynyl, cycloalkyl, substitutedcycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic and substituted heterocyclic are as defined herein.

“Oxysulfonylamino” refers to the groups —OSO₂NH₂, —OSO₂NRR,—OSO₂NR-alkyl, —OSO₂NR-substituted alkyl, —OSO₂NR-alkenyl,—OSO₂NR-substituted alkenyl, —OSO₂NR-alkynyl, —OSO₂NR-substitutedalkynyl, —OSO₂NR-cycloalkyl, —OSO₂NR-substituted cycloalkyl,—OSO₂NR-aryl, —OSO₂NR-substituted aryl, —OSO₂NR-heteroaryl,—OSO₂NR-substituted heteroaryl, —OSO₂NR-heterocyclic, and—OSO₂NR-substituted heterocyclic where R is hydrogen, alkyl or whereeach R is joined to form, together with the nitrogen atom a heterocyclicor substituted heterocyclic ring and wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Oxythiocarbonylamino” refers to the groups —OC(S)NH₂, —OC(S)NRR,—OC(S)NR-alkyl, —OC(S)NR-substituted alkyl, —OC(S)NR-alkenyl,—OC(S)NR-substituted alkenyl, —OC(S)NR-alkynyl, —OC(S)NR-substitutedalkynyl, —OC(S)NR-cycloalkyl, —OC(S)NR-substituted cycloalkyl,—OC(S)NR-aryl, —OC(S)NR-substituted aryl, —OC(S)NR-heteroaryl,—OC(S)NR-substituted heteroaryl, —OC(S)NR-heterocyclic, and—OC(S)NR-substituted heterocyclic where R is hydrogen, alkyl or whereeach R is joined to form together with the nitrogen atom a heterocyclicor substituted heterocyclic ring and wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Thioalkyl” refers to the groups —S-alkyl.

“Substituted thioalkyl” refers to the group —S-substituted alkyl.

“Thioamidino” refers to the group RSC(═NH)— where R is hydrogen oralkyl.

“Thioaryl” refers to the group —S-aryl and “substituted thioaryl” refersto the group —S-substituted aryl.

“Thiocarbonylamino” refers to the group —C(S)NRR where each R isselected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl,substituted aryl, cycloalkyl, substituted cycloalkyl, heteroaryl,substituted heteroaryl, heterocyclic, substituted heterocyclic and whereeach R is joined to form, together with the nitrogen atom a heterocyclicor substituted heterocyclic ring wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Thiocycloalkyl” refers to the groups —S-cycloalkyl.

“Substituted thiocycloalkyl” refers to the group —S-substitutedcycloalkyl.

“Thioheteroaryl” refers to the group —S-heteroaryl and “substitutedthioheteroaryl” refers to the group —S-substituted heteroaryl.

“Thioheterocyclic” refers to the group —S-heterocyclic and “substitutedthiobeterocyclic” refers to the-group —S-substituted heterocyclic.

“Thiol” refers to the group —SH.

“Optionally substituted” means that the recited group may beunsubstituted or the recited group may be substituted.

“Pharmaceutically acceptable carrier” means a carrier that is useful inpreparing a pharmaceutical composition or formulation that is generallysafe, non-toxic and neither biologically nor otherwise undesirable, andincludes a carrier that is acceptable for veterinary use as well ashuman pharmaceutical use. A pharmaceutically acceptable carrier orexcipient as used in the specification and claims includes both one ormore than one of such carriers.

“Pharmaceutically-acceptable cation” refers to the cation of apharmaceutically-acceptable salt.

“Pharmaceutically acceptable salt” refers to salts which retain thebiological effectiveness and properties of the compounds of thisinvention and which are not biologically or otherwise undesirable.Pharmaceutically acceptable salts refer to pharmaceutically acceptablesalts of the compounds, which salts are are derived from a variety oforganic and inorganic counter ions well known in the art and include, byway of example only, sodium, potassium, calcium, magnesium, ammonium,tetraalkylammonium, and the like; and when the molecule contains a basicfunctionality, salts of organic or inorganic acids, such ashydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate,oxalate and the like.

Pharmaceutically-acceptable base addition salts can be prepared frominorganic and organic bases. Salts derived from inorganic bases, includeby way of example only, sodium, potassium, lithium, ammonium, calciumand magnesium salts. Salts derived from organic bases include, but arenot limited to, salts of primary, secondary and tertiary amines, such asalkyl amines, dialkyl amines, trialkyl amines, substituted alkyl amines,di(substituted alkyl) amines, tri(substituted alkyl) amines, alkenylamines, dialkenyl amines, trialkenyl amines, substituted alkenyl amines,di(substituted alkenyl) amines, tri(substituted alkenyl) amines,cycloalkyl amines, di(cycloalkyl) amines, tri(cycloalkyl) amines,substituted cycloalkyl amines, disubstituted cycloalkyl amine,trisubstituted cycloalkyl amines, cycloalkenyl amines, di(cycloalkenyl)amines, tri(cycloalkenyl) amines, substituted cycloalkenyl amines,disubstituted cycloalkenyl amine, trisubstituted cycloalkenyl amines,aryl amines, diaryl amines, triaryl amines, heteroaryl amines,diheteroaryl amines, triheteroaryl amines, heterocyclic amines,diheterocyclic amines, triheterocyclic amines, mixed di- and tri-amineswhere at least two of the substituents on the amine are different andare selected from the group consisting of alkyl, substituted alkyl,alkenyl, substituted alkenyl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl, heterocyclic,and the like. Also included are amines where the two or threesubstituents, together with the amino nitrogen, form a heterocyclic orheteroaryl group.

Examples of suitable amines include, by way of example only,isopropylamine, trimethyl amine, diethyl amine, tri(iso-propyl) amine,tri(n-propyl) amine, ethanolamine, 2-dimethylaminoethanol, tromethamine,lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline,betaine, ethylenediamine, glucosamine, N-alkylglucamines, theobromine,purines, piperazine, piperidine, morpholine, N-ethylpiperidine, and thelike. It should also be understood that other carboxylic acidderivatives would be useful in the practice of this invention, forexample, carboxylic acid amides, including carboxamides, lower alkylcarboxamides, dialkyl carboxamides, and the like.

Pharmaceutically acceptable acid addition salts may be prepared frominorganic and organic acids. Salts derived from inorganic acids includehydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, and the like. Salts derived from organic acids includeacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid,malic acid, malonic acid, succinic acid, maleic acid, fumaric acid,tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, p-toluene-sulfonic acid,salicylic acid, and the like.

A “therapeutically effective amount” means the amount of a compound orantibody that, when administered to a mammal for treating a disease, issufficient to effect such treatment for the disease. The“therapeutically effective amount” will vary depending on the compound,the disease and its severity and the age, weight, etc., of the mammal tobe treated.

As used herein, the following abbreviations have the following meanings.If an abbreviation is not defined, it has its generally acceptedmeaning.

-   -   AC=acid ceramidase    -   CAN=acetonitrile    -   AcOH=acetic acid    -   ACTH=adrenocorticotropic hormone    -   ADEM=acute disseminated encephalomyelitis    -   ANA=Anti-nuclear antibodies    -   aq or aq.=aqueous    -   AUC=Area under the curve    -   BBB=blood brain barrier    -   bd=broad doublet    -   bm=broad multiplet    -   Bn=benzyl    -   Boc=tert-butoxycarbonyl    -   Boc₂O=di-tert-butyl dicarbonate    -   BOP=benzotriazol-1-yloxy-tris(dimethylamino)phosphonium        hexafluorophosphate    -   bs=broad singlet    -   BSA=bovine serum albumin    -   C=constant region of an immunoglobulin    -   Cbz=carbobenzyloxy    -   cDNA=complementary deoxyribnucleic acid    -   CDR=complementarity determining region    -   CDR1=complementarity determining region 1    -   CDR2=complementarity determining region 2    -   CDR3=complementarity determining region 3    -   CFA=complete Freund's adjuvant    -   CHCl₃=chloroform    -   CH₂Cl₂=dichloromethane    -   CIDP=chronic immune demyelinating polyneuropathy    -   CNS=central nervous system    -   (COCl)₂=oxalyl chloride    -   COX-2=cyclooxygenase-2    -   CRP=C-Reactive Protein    -   CS=Cockayne's syndrome    -   CSF=colony stimulating factor    -   d=doublet    -   DBU=1,8-diazabicyclo[5.4.0]undec-7-ene    -   DCC=1,3-dicyclohexylcarbodiimide    -   dd=doublet of doublets    -   DMAP=4-N,N-dimethylaminopyridine ethylcarbodiimide hydrochloride    -   DME=ethylene glycol dimethyl ether    -   DMF=N,N-dimethylformamide    -   DMSO=dimethylsulfoxide    -   DNA=deoxyribonucleic acid    -   dt=doublet of triplets    -   EAE=experimental autoimmune encephalomyelitis    -   EBNA2=Epstein-Barr virus nuclear antigen 2    -   ECM=extracellular matrix    -   EDC=1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride    -   EDTA=Ethylenediamine tetraacetic acid    -   ELAMS=endothelial adhesion molecules    -   EM=electron microscopy    -   Et₃N=triethylamine    -   Et₂O=diethyl ether    -   EtOAc=ethyl acetate    -   EtOH=ethanol    -   eq or eq.=equivalent    -   FACS=Fluorescence activated Cell Sorter    -   FITC=Fluorescein isothiocyanate    -   Fmoc=N-(9-fluorenylmethoxycarbonyl)    -   FmocONSu=N-(9-fluorenylmethoxycarbonyl)-succinimide    -   FR=framework region    -   FRI=framework region 1    -   FR2=framework region 2    -   FR3=framework region 3    -   g=grams    -   GM-CSF=granulocyte monocyte colony stimulating factor    -   h or hr=hour    -   H=heavy chain of an immunoglobulin    -   HAMA=human anti-mouse antibody    -   HB or Hb=hemoglobin    -   HBr=hydrobromic acid    -   HBSS=Hank's Balanced Saline Solution    -   HCl=hydrochloric acid    -   Hct=hematocrit, or measurement of packed red blood cells        obtained by centrifugation in a volume of a blood sample    -   H-E=hematoxylin-eosin    -   HEPES=4-(2-hydroxyethyl)-1-piperazine-ethanesulfonic acid    -   hex A=hexoaminidase A    -   HIC=Hydrophobic interaction chromatography    -   HIG=human immunoglobulin    -   H₂O=water    -   HOBT=1-hydroxybenzotriazole hydrate    -   HUVEC=human umbilical vascular endothelial cells    -   IgG Fc=a binding domain of the immunoglobulin    -   i.p.=intraperitoneal    -   ICAM-1=intercellular adhesion molecule 1    -   Ig=immunoglobulin    -   IgG=immunoglobulin G    -   IgM=immunoglobulin M    -   IL=interleukin    -   IL-1=interleukin-1    -   IL-2=interleukin-2    -   IL-8=interleukin-8    -   IBD=inflammatory bowel disease    -   IBDQ=inflammatory bowel disease questionairre    -   ITT=Intention-to-treat (including all subjects randomized,        regardless of whether dosed)    -   K₂CO₃=potassium carbonate    -   kg=kilogram    -   L=liter    -   LCMS=Liquid chromatography Mass Spectroscopy    -   LFA-1=lymphocyte function-related antigen 1—(also known as β₂        integrin, CD11a/CD18 and α_(L)β₂)    -   m=multiplet (when used with NMR data)    -   M=Molar    -   MAbs1=monoclonal antibodies    -   Mac-1=α_(M)β₂ integrin (also known as CD11b/CD18)    -   MAdCAM-1=mucosal addressin cell adhesion molecule    -   MALDI/TOF MS=matrix-assisted laser desorption        ionization/time-of-flight mass spectrometry    -   MALDI/TOF MS=matrix-assisted laser desorption        ionization/time-of-flight mass spectrometry    -   MBP=myelin basic protein    -   MCH=Mean Corpusular Hemoglobin; Hb/RBC    -   MCHC=mean corpuscular hemoglobin count expressed as a        percentage; Hb/Hct.    -   MCP-1=monocyte chemotactic protein 1    -   MCV=mean corpuscular volume; the avg. volume of erythrocytes,        conventionally expressed in cubic micrometers per red cell.    -   MeOH=methanol    -   MES=2-(N-morpholino)ethanesulfonic acid    -   mg=milligram    -   MgSO₄=magnesium sulfate    -   min.=minute    -   MIP-1α=macrophage inflammatory protein 1 alpha    -   MIP-1β=macrophage inflammatory protein 1 beta    -   mL=milliliter    -   MLD=metachromatic leukodystrophy    -   mm=millimeter    -   mM=millimolar    -   MOG=myelin-oligodendrocyte glycoprotein    -   mol=moles    -   mmol=millimol    -   mp=melting point    -   mpk=milligrams per killogram    -   MS=multiple sclerosis    -   N=normal    -   NaCl=sodium chloride    -   Na₂CO₃=sodium carbonate    -   NaHCO₃=sodium bicarbonate    -   NaOEt=sodium ethoxide    -   NaOH=sodium hydroxide    -   ng=nanograms    -   NH₄Cl=ammonium chloride    -   NMM=N-methylmorpholine    -   NSAID=nonsteroidal anti-inflammatory    -   OtBu=tert-butoxy    -   PBS++=Phosphate buffered saline    -   PCR=polymerase chain reaction    -   PEG=polyethylene glycol    -   Phe=L-phenylalanine    -   PKU=phenylketonuria    -   PLP=proteolipid protein    -   POEMS=polyneuropathy organomegaly endocrinopathy, M-protein and        skin changes    -   PMSF=phenylmethylsulfonylfluoride    -   Pro=L-proline    -   PRP=prion related protein    -   psi=pounds per square inch    -   PtO₂=platinum oxide    -   q=quartet    -   quint.=quintet    -   q.s. or Q.S.=bring to volume    -   RA=rheumatoid arthritis    -   RANTES=regulated upon activation, normal T-cell expressed and        secreted chemokine (also known as small inducible cytokine A5)    -   RBC=red blood cell count    -   Rfs or R_(f)=retention factor    -   RNA=ribonucleic acid    -   rpm=rotations per minute    -   rt or RT=room temperature    -   RT-PCR=reverse transcription polymerase chain reaction    -   s=singlet    -   SAE=Serious adverse event    -   SAMIs=selective adhesion molecule inhibitors    -   sat or sat.=saturated    -   scFv=single chain Fv fragment    -   SCR=solochrome-R-cyanlin    -   SDS=sodium dodecyl sulfate    -   SDS-PAGE=sodium dodecyl sulfate polyacrylamide gel        electrophoresis    -   t=triplet    -   t-BuOH=tert-butanol    -   TFA=trifluoroacetic acid    -   TGF-β=tumor growth factor beta    -   THF=tetrahydrofuran    -   TLC or tlc=thin layer chromatography    -   TNF=tumor necrosis factor    -   TNF-α=tumor necrosis factor alpha    -   TNF-β=tumor necrosis factor beta    -   Ts=tosyl    -   TsCl=tosyl chloride    -   TsOH=tosylate    -   TYR=tyrosine    -   μg=microgram    -   μL=microliter    -   μM=micromolar    -   μm=microns    -   UV=ultraviolet    -   VCAM-1=vascular cell adhesion molecule 1    -   V_(H)=heavy chain of the variable domain    -   V_(L)=light chain of the variable domain    -   VLA-4=very late antigen 4 (also known as alpha-4 beta-1, α₄β₁)    -   V_(t)=Total volume    -   WBC=White Blood Cells    -   w/w=weight to weight    -   w/v=weight to volume    -   v/v=volume to volume    -   φ=phenyl

GENERAL ASPECTS OF THE INVENTION

The present invention is based on the surprising result that combinationtherapies of methotrexate and antibodies to alpha-4 integrin, such asAntegren™, as well as combination therapies of methotrexate and smallmolecule alpha-4 antagonists effectively suppress the deleteriouseffects of RA.

Methotrexate

Methotrexate (Amethopterin®, Mexate®, Folex® and Rheumatrex®) interfereswith the production and maintenance of DNA. It is not understood exactlyhow methotrexate works in rheumatoid arthritis, but reduces inflammationand slow worsening of the disease. Methotrexate is considered adisease-modifying antirheumatic drug (DMARD). It is effective in theearly stages of rheumatoid arthritis to prevent disease progression,especially in combination with other medications.

Methotrexate is effective in relieving joint inflammation, slowingdisease progression, and preventing disability by delaying jointdestruction. Patients with rheumatoid arthritis may be more likely tocontinue treatment with methotrexate than with other DMARDs because offavorable results and tolerable side effects. Physicians often recommendthat methotrexate be used with one or more combination therapy.

Combination therapy may allow for lower doses of an individual drug tobe used, which may reduce the risk of adverse effects that can occurwith higher doses. Methotrexate combined with hydroxychloroquine andsulfasalazine may be more effective that methotrexate alone.

Whatever may be the actual cause, there is no cure for RA, and althoughthe disease is not fatal, disease complications and symptoms may persistthroughout an individual's lifetime, and may even shorten survival by afew years. Affected joints may become deformed, and the performance ofeven ordinary tasks may be very difficult or impossible.

Notwithstanding what has been previously reported in the literature, newcompounds, compositions and methods for using these compounds andcompositions to inhibit rheumatoid arthritis are needed.

In a general sense, the method of the invention does not involve anyparticular mode of administration, since the mode of administration isdependent upon the form of the active agent and the formulationdeveloped to administer the active agent. Modes of administrationinclude, but are not limited to, oral, parenteral (e.g., subcutaneous,subdural, intravenous, intramuscular, intrathecal, intraperitoneal,intracerebral, intraarterial, or intralesional routes ofadministration), topical, localized (e.g., surgical application orsurgical suppository), rectal, and pulmonary (e.g., aerosols,inhalation, or powder). The route of administration would be based onthe composition being administered (e.g., immunoglobulin beingadministered intravenously versus small compound being administeredorally), tissue targeting (e.g., intrathecal administration to targetthe site of a spinal cord injury), and the like, as would be known tothe artisan of ordinary skill.

Additionally, the anti-alpha-4 agents (e.g., anti-alpha-4-antibodies,small compound alpha-4-integrin antagonists, and the like) can becombined with other compounds or compostions used to treat, ameliorateor palliate symptoms associated with rheumatoid arthritis. Furthermore,the compounds disclosed herein can be administered alone or incombination with other agents, such as alpha-4 integrin inhibitors,including anti-alpha-4 integrin antibodies and immunologically activefragments thereof (e.g., natalizumab). When administered in combination,the small compound alpha-4-integrin antagonists may be administered inthe same formulation as these other compounds or compositions, or in aseparate formulation. When administered in combination, theanti-alpha-4-antibodies are generally administered in a separateformulation than the small compound alpha-4-integrin antagonists, othercompounds, and compositions. When administered in combinations, theanti-alpha-4 agents may be administered prior to, following, orconcurrently with the other compounds and compositions used to treat,ameliorate, or palliate symptoms. The invention relates to introducingrelatively constant amounts of an active agent to a patient'scirculatory system over a period of months or years. This chronicintroduction of an agent that selectively-binds to alpha-4 integrin or adimer comprising alpha-4 integrin (e.g. i-alpha-4 beta-1) results insuppression of pathological inflammation being maintained at a constantlevel over a period of time. By maintaining therapeutic levels of anactive agent for a period of time, pathological inflammation can bechronically suppressed in the patient.

Compounds of Formulae I and II

In one aspect, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compounds ofFormulae I, IA, IB, IC, and II.

In one aspect, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compoundsdefined by Formula I below. These compounds have a binding affinity toVLA-4 as expressed by an IC₅₀ of about 15 μM or less (measured asdescribed in Example A below):

wherein

-   -   R¹ is selected from the group consisting of alkyl, substituted        alkyl, aryl, substituted aryl, cycloalkyl, substituted        cycloalkyl, heterocyclic, substituted heterocylic, heteroaryl        and substituted heteroaryl;    -   R² is selected from the group consisting of hydrogen, alkyl,        cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted        cycloalkenyl, heterocyclic, substituted heterocyclic,        substituted alkyl, aryl, substituted aryl, heteroaryl,        substituted heteroaryl, and R¹ and R² together with the nitrogen        atom bound to R² and the SO₂ group bound to R¹ can form a        heterocyclic or a substituted heterocyclic group;    -   R³ is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl, substituted heteroaryl,        heterocyclic, substituted heterocyclic and, when R² does not        form a heterocyclic group with R¹, R² and R³ together with the        nitrogen atom bound to R² and the carbon atom bound to R³ can        form a heterocyclic or a substituted heterocyclic group;    -   R⁵ is —(CH₂)_(x)—Ar—R⁵ where R^(5′) is selected from the group        consisting of —O—Z—NR⁸R^(8′) and —O—Z—R^(8″) wherein R⁸ and        R^(8′) are independently selected from the group consisting of        hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted        cycloalkyl, heterocyclic, substituted heterocyclic, and where R⁸        and R^(8′) are joined to form a heterocycle or a substituted        heterocycle, R^(8″) is selected from the group consisting of        heterocycle and substituted heterocycle, and Z is selected from        the group consisting of —C(O)— and —SO₂—;    -   Ar is aryl, heteroaryl, substituted aryl or substituted        heteroaryl;    -   x is an integer of from 1 to 4;    -   Q is —C(X)NR⁷— wherein R⁷ is selected from the group consisting        of hydrogen and alkyl; and X is selected from the group        consisting of oxygen and sulfur;    -   and pharmaceutically acceptable salts thereof.

In another embodiment, the compounds can be provided as prodrugs whichconvert (e.g., hydrolyze, metabolize, etc.) in vivo to a compound ofFormula I above. In a preferred example of such an embodiment, thecarboxylic acid group of the compound of Formula I is modified into agroup which, in vivo, will convert to a carboxylic acid group (includingsalts thereof). In a particularly preferred embodiment, such prodrugsare represented by compounds of Formula IA:

wherein:

-   -   R¹ is selected from the group consisting of alkyl, substituted        alkyl, aryl, substituted aryl, cycloalkyl, substituted        cycloalkyl, heterocyclic, substituted heterocylic, heteroaryl        and substituted heteroaryl;    -   R² is selected from the group consisting of hydrogen, alkyl,        cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted        cycloalkenyl, heterocyclic, substituted heterocyclic,        substituted alkyl, aryl, substituted aryl, heteroaryl,        substituted heteroaryl, and R¹ and R² together with the nitrogen        atom bound to R² and the SO₂ group bound to R¹ can form a        heterocyclic or a substituted heterocyclic group;    -   R³ is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl, substituted heteroaryl,        heterocyclic, substituted heterocyclic and, when R² does not        form a heterocyclic group with R¹, R² and R³ together with the        nitrogen atom bound to R² and the carbon atom bound to R³ can        form a heterocyclic or a substituted heterocyclic group;    -   R⁵ is —(CH₂)_(x)—Ar—R^(5′) where R^(5′) is selected from the        group consisting of —O—Z—NR⁸R^(8′) and —O—Z—R^(8″) wherein R⁸        and R^(8′) are independently selected from the group consisting        of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted        cycloalkyl, heterocyclic, substituted heterocyclic, and where R⁸        and R^(8′) are joined to form a heterocycle or a substituted        heterocycle, R^(8″) is selected from the group consisting of        heterocycle and substituted heterocycle, and Z is selected from        the group consisting of—C(O)— and —SO₂—;    -   Ar is aryl, heteroaryl, substituted aryl or substituted        heteroaryl;    -   x is an integer of from 1 to 4;    -   R⁶ is selected from the group consisting of        2,4-dioxo-tetrahydrofuran-3-yl (3,4-enol), amino, alkoxy,        substituted alkoxy, cycloalkoxy, substituted cycloalkoxy,        —O—(N-succinimidyl), —NH-adamantyl, —O-cholest-5-en-3-O-yl,        —NHOY where Y is hydrogen, alkyl, substituted alkyl, aryl, and        substituted aryl, —NH(CH₂)_(p)COOY where p is an integer of from        1 to 8 and Y is as defined above, —OCH₂NR⁹R¹⁰ where R⁹ is        selected from the group consisting of —C(O)-aryl and        —C(O)-substituted aryl and R¹⁰ is selected from the group        consisting of hydrogen and —CH₂COOR¹¹ where R¹¹ is alkyl, and        —NHSO₂Z′ where Z′ is alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, aryl, substituted aryl, heteroaryl,        substituted heteroaryl, heterocyclic and substituted        heterocyclic;    -   Q is —C(X)NR⁷— wherein R⁷ is selected from the group consisting        of hydrogen and alkyl; and X is selected from the group        consisting of oxygen and sulfur;    -   and pharmaceutically acceptable salts thereof    -   with the following provisos    -   (A) when R¹ and R² together with the SO₂ group pendent to R¹ and        the nitrogen pendent to R² form a saccharin-2-yl group, R³ is        —CH₃, R⁵ is p-[(CH₃)₂NC(O)-]benzyl and Q is —C(O)NH— then R⁶ is        not —OC(CH₃)₃;    -   (B) when R¹ is p-methylphenyl, R² and R³ together with the        nitrogen atom pendent to R² and the carbon atom pendent to R³        form a pyrrodinyl ring derived from D-proline; R⁵ is        p-[(4-methylpiperazin-1-yl)NC(O)O-]benzyl derived from        D-phenylalanine and Q is —C(O)NH— then R⁶ is not —OC(CH₃)₃;    -   (C) when R¹ is pyrimidin-2-yl, R² and R³ together with the        nitrogen atom bound to R² and the carbon atom bound to R³ form a        pyrrolidinyl ring, R⁵ is p-[(CH₃)₂NC(O)O-]benzyl and Q is        —C(O)NH— then R is not —OC(CH₃)₃; and    -   (D) when R¹ is p-methylphenyl, R² and R³ together with the        nitrogen atom pendent to R² and the carbon atom pendent to R³        form a (2S)-piperazin-2-carbonyl ring; R⁵ is        p-[(CH₃)₂NC(O)O-]benzyl and Q is —C(O)NH— then R⁶ is not        —OC(CH₃)₃.

Further description of the compounds of the above Formulae I and IA andprocedures and reaction conditions for preparing these compounds aredescribed in U.S. Ser. No. 09/126,958 (filed Jul. 31, 1998 and issued asU.S. Pat. No. 6,489,300), herein incorporated by reference in itsentirety.

Preferably, in the compounds of Formulae I and IA above, R¹ is selectedfrom the group consisting of alkyl, substituted alkyl, aryl, substitutedaryl, heterocyclic, substituted heterocylic, heteroaryl and substitutedheteroaryl. More preferably R¹ is selected from the group consisting ofaryl, substituted aryl, heteroaryl and substituted heteroaryl.

Preferably R¹, in the compounds of Formulae I and IA above is selectedfrom the group consisting of phenyl, 4-methylphenyl, 4-t-butylphenyl,2,4,6-trimethylphenyl, 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl,2,4-difluorophenyl, 3,4-difluorophenyl, 3,5-difluorophenyl,2-chlorophenyl, 3-chlorophenyl, 4-chlorophenyl, 3,4-dichlorophenyl,3,5-dichlorophenyl, 3-chloro-4-fluorophenyl, 4-bromophenyl,2-methoxyphenyl, 3-methoxyphenyl, 4-methoxyphenyl, 3,4-dimethoxyphenyl,4-t-butoxyphenyl, 4-(3′-dimethylamino-n-propoxy)-phenyl,2-carboxyphenyl, 2-(methoxycarbonyl)phenyl, 4-(H₂NC(O)-)phenyl,4-(H₂NC(S)-)phenyl, 4-cyanophenyl, 4-trifluoromethylphenyl,4-trifluoromethoxyphenyl, 3,5-di-(trifluoromethyl)phenyl, 4-nitrophenyl,4-aminophenyl, 4-(CH₃C(O)NH-) phenyl, 4-(PhNHC(O)NH-)phenyl,4-amidinophenyl, 4-methylamidinophenyl, 4-(CH₃SC(═NH)-)phenyl,4-chloro-3-(H₂NS(O)₂-)phenyl, 1-naphthyl, 2-naphthyl, pyridin-2-yl,pyridin-3-yl, pyrimidin-2-yl, quinolin-8-yl,2-(trifluoroacetyl)-1,2,3,4-tetrahydroisoquinolin-7-yl, morpholin-4-yl,2-thienyl, 5-chloro-2-thienyl, 2,5-dichloro-4-thienyl,1-N-methylimidazol-4-yl, 1-N-methylpyrazol-3-yl, 1-N-methylpyrazol-4-yl,1-N-butylpyrazol-4-yl, 1-N-methyl-3-methyl-5-chloropyrazol-4-yl,1-N-methyl-5-methyl-3-chloropyrazol-4-yl, 2-thiazolyl and5-methyl-1,3,4-thiadiazol-2-yl.

Preferably, R², in the compounds of Formulae I and IA above is selectedfrom the group consisting of methyl, benzyl, —(CH₂)₂-2-thienyl, and—(CH₂)₂-φ.

In one preferred embodiment, R² and R³, in the compounds of Formulae Iand IA above together with the nitrogen atom bound to the R² substituentand the carbon bound to the R³ substituent form a heterocyclic group ora substituted heterocyclic group of 4 to 6 ring atoms having 1 to 2heteroatoms in the ring selected from the group consisting of nitrogen,oxygen and sulfur which ring is optionally substituted with 1 to 2substituents selected from the group consisting of fluoro, methyl,hydroxy, oxo (═O), amino, phenyl, thiophenyl, thiobenzyl,(thiomorpholin-4-yl)C(O)O—, CH₃S(O)₂— and CH₃S(O)₂O—, or can be fused toanother ring such as a phenyl or cycloalkyl ring to provide for a fusedring heterocycle of from 10 to 14 ring atoms having 1 to 2 heteroatomsin the ring selected from the group consisting of nitrogen, oxygen andsulfur. Such heterocyclic rings include azetidinyl (e.g., L-azetidinyl),thiazolidinyl (e.g., L-thiazolidinyl), piperidinyl (e.g.,L-piperidinyl), piperazinyl (e.g., L-piperazinyl), dihydroindolyl (e.g.,L-2,3-dihydroindol-2-yl), tetrahydroquinolinyl (e.g.,L-1,2,3,4-tetrahydroquinolin-2-yl), thiomorpholinyl (e.g.,L-thiomorpholin-3-yl), pyrrolidinyl (e.g., L-pyrrolidinyl), substitutedpyrrolidinyl such as 4-hydroxypyrrolidinyl (e.g., 4-α-(orβ-)hydroxy-L-pyrrolidinyl), 4-oxopyrrolidinyl (e.g.,4-oxo-L-pyrolidinyl), 4-fluoropyrrolidinyl (e.g., 4-α-(orβ-)fluoro-L-pyrrolidinyl), 4,4-difluoropyrrolidinyl (e.g.,4,4-difluoro-L-pyrrolidinyl), 4-(thiomorpholin-4-ylC(O)O-)pyrrolidinyl(e.g., 4-α-(or β-)-(thiomorpholin-4-ylC(O)O—)-L-pyrrolidinyl,4-(CH₃S(O)₂O-)pyrrolidinyl (e.g., 4-α-(orβ-)(CH₃S(O)₂O-)-L-pyrrolidinyl, 3-phenylpyrrolidinyl (e.g., 3-α-(orβ-)phenyl-L-pyrrolidinyl), 3-thiophenylpyrrolidinyl (e.g, 3-α-(orβ-)-thiophenyl-L-pyrrolidinyl), 4-aminopyrrolidinyl (e.g., 4-α-(orβ-)amino-L-pyrrolidinyl), 3-methoxypyrrolidinyl (e.g., 3-α-(orβ-)methoxy-L-pyrrolidinyl), 4,4-dimethylpyrrolidinyl, substitutedpiperazinyl such as 4-N-Cbz-piperazinyl and 4-(CH₃S(O)₂-)piperazinyl,substituted thiazolidinyl such as 5,5-dimethylthiazolindin-4-yl,1,1-dioxo-thiazolidinyl (e.g., L-1,1-dioxo-thiazolidin-2-yl),substituted 1,1-dioxo-thiazolidinyl such asL-1,1-dioxo-5,5-dimethylthiazolidin-2-yl, 1,1-dioxothiomorpholinyl(e.g., L-1,1-dioxo-thiomorpholin-3-yl) and the like.

Q, in the compounds of Formulae I and IA above, is preferably —C(O)NH—or —C(S)NH—.

In the compounds of Formulae I and IA above, Ar is preferably aryl orsubstituted aryl and, even more preferably, is phenyl or substitutedphenyl. Preferably, x is 1.

In the compounds of Formulae I and IA above, R⁵ is preferably selectedfrom all possible isomers arising by substitution with the followinggroups:

-   -   3-[(CH₃)₂NC(O)O-]benzyl,    -   4-[(CH₃)₂NC(O)O-]benzyl,    -   4-[(piperidin-1′-yl)C(O)O-]benzyl,    -   4-[(piperidin-4′-yl)C(O)O-]benzyl,    -   4-[(1′-methylpiperidin-4′-yl)C(O)O-]benzyl,    -   4-[(4′-hydroxypiperidin-1′-yl)C(O)O-]benzyl,    -   4-[(4′-formyloxypiperidin-1′-yl)C(O)O-]benzyl,    -   4-[(4′-ethoxycarbonylpiperidin-1′-yl)C(O)O-]benzyl,    -   4-[(4′-carboxylpiperidin-1′-yl)C(O)O-]benzyl,    -   4-[(3 ′-hydroxymethylpiperidin-1′-yl)C(O)O-]benzyl,    -   4-[(4′-hydroxymethylpiperidin-1′-yl)C(O)O-]benzyl,    -   4-[(4′-phenyl-1′-Boc-piperidin-4′-yl)—C(O)O-]benzyl,    -   4-[(4′-piperidon-1′-yl ethylene ketal)C(O)O-]benzyl,    -   4-[(piperazin-4′-yl)—C(O)O-]benzyl,    -   4-[(1′-Boc-piperazin-4′-yl)—C(O)O-]benzyl,    -   4-[4′-meth-ylpiperazin-yl)C(O)O-]benzyl,    -   4-[(4′-methylhomopiperazin-1′-yl)C(O)O-]benzyl,    -   4-[(4′-(2-hydroxyethyl)piperazin-1′-yl)C(O)O-]benzyl,    -   4-[(4′-phenylpiperazin-1′-yl)C(O)O-]benzyl,    -   4-[(4′-(pyridin-2-yl)piperazin-1′-yl)C(O)O-]benzyl,    -   4-[(4′-(4-trifluoromethylpyridin-2-yl)piperazin-1′-yl)C(O)O-]benzyl,    -   4-[(4′-(pyrimidin-2-yl)piperazin-1′-yl)C(O)O-]benzyl,    -   4-[(4′-acetylpiperazin-1′-yl)C(O)O-]benzyl,    -   4-[(4′-(phenylC(O)-)piperazin-1′-yl)C(O)O-]benzyl,    -   4-[(4′-(pyridin-4-ylC(O)-)piperazin-1′-yl)C(O)O-]benzyl,    -   4-[(4′-(phenylNHC(O)-)piperazin-1′-yl)C(O)O-]benzyl,    -   4-[(4′-(phenylNHC(S)-)piperazin-1′-yl)C(O)O-]benzyl,    -   4-[(4′-methanesulfonylpiperazin-1′-yl—C(O)O-)benzyl,    -   4-[(4′-trifluoromethanesulfonylpiperazin-1′-yl—C(O)O-)benzyl,    -   4-[(morpholin-4′-yl)C(O)O-]benzyl,    -   3-nitro-4-[(morpholin-4′-yl)—C(O)O-]benzyl,    -   4-[(thiomorpholin-4 ′-yl)C(O)O-]benzyl,    -   4-[(thiomorpholin-4′-yl sulfone)—C(O)O-]benzyl (alternative        nomenclature 4-[(1,1-dioxothiomorpholin-4-yl)—C(O)O-]benzyl),    -   4-[(pyrrolidin-1′-yl)C(O)O-]benzyl,    -   4-[(2′-methylpyrrolidin-1′-yl)C(O)O-]benzyl,    -   4-[(2′-(methoxycarbonyl)pyrrolidin-1′-yl)C(O)O-]benzyl,    -   4-[(2′-(hydroxymethyl)pyrrolidin-1′-yl)C(O)O-]benzyl,    -   4-[(2′-(N,N-dimethylamino)ethyl)(CH₃)NC(O)O-]benzyl,    -   4-[(2′-(N-methyl-N-toluene-4-sulfonylamino)ethyl)(CH₃)NC(O)O-]benzyl,    -   4-[(2′-(morpholin-4′-yl)ethyl)(CH₃)NC(O)O-]benzyl,    -   4-[(2′-(hydroxy)ethyl)(CH₃)NC(O)O-]benzyl,    -   4-[bis(2′-(hydroxy)ethyl)NC(O)O-]benzyl,    -   4-[(2′-(formyloxy)ethyl)(CH₃)NC(O)O-]benzyl,    -   4-[(CH₃OC(O)CH₂)HNC(O)O-]benzyl,    -   4-[2′-(phenylNHC(O)O-)ethyl-]HNC(O)O-]benzyl,    -   3-chloro-4-[(CH₃)₂NC(O)O-]benzyl,    -   3-chloro-4-[(4′-methylpiperazin-1′-yl)C(O)O-]benzyl,    -   3-chloro-4-[(4′-(pyridin-2-yl)piperazin-1′-yl)C(O)O-]benzyl,    -   3-chloro-4-[(thiomorpholin-4′-yl)C(O)O-]benzyl, and    -   3-fluoro-4-[(CH₃)₂NC(O)O-]benzyl.

In the compounds of Formula IA, R⁶ is preferably2,4-dioxo-tetrahydrofuran-3-yl (3,4-enol), methoxy, ethoxy, n-propoxy,isopropoxy, n-butoxy, t-butoxy, cyclopentoxy, cyclopropylmethoxy,neopentoxy, 2-α-isopropyl-4-β-methylcyclohexoxy,2-β-isopropyl-4-β-methylcyclohexoxy, 2-methoxyphenoxy,2-(morpholin-4-yl)ethoxy, —O(CH₂CH₂O)₂CH₃, 2-(phenoxy)ethoxy,—OCH₂C(CH₃)₂NHBoc, —NH₂, benzyloxy, —NHCH₂COOH, —NHCH₂CH₂COOH,—NH-adamantyl, —NHSO₂-p—CH₃-φ, —NHCH₂CH₂COOCH₂CH₃, —NHOY′ where Y′ ishydrogen, methyl, iso-propyl or benzyl, O—(N-succinimidyl),—O-cholest-5-en-3-β-yl, —OCH₂—OC(O)C(CH₃)₃, —O(CH₂)_(z)NHC(O)W where zis 1 or 2 and W is selected from the group consisting of pyrid-3-yl,N-methylpyridyl, and N-methyl-1,4-dihydro-pyrid-3-yl, —NR″C(O)—R′ whereR′ s aryl, heteroaryl or heterocyclic and R″ is hydrogen or—CH₂C(O)OCH₂CH₃.

Even more preferably, R⁶ in the compounds of Formula IA is selected fromthe group consisting of methoxy, ethoxy, n-propoxy, isopropoxy,n-butoxy, t-butoxy, cyclopentoxy, cyclopropylmethoxy, neopentoxy,2-α-isopropyl-4-β-methylcyclohexoxy,2-β-isopropyl-4-β-methylcyclohexoxy, 2-methoxyphenoxy,2-(morpholin-4-yl)ethoxy, —O(CH₂CH₂O)₂CH₃, 2-(phenoxy)ethoxy,—OCH₂C(CH₃)₂NHBoc, and benzyloxy.

Preferred compounds within the scope of Formulae I and IA above includeby way of example:

-   -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        ethyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        ethyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-methyl-piperazin-1-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        n-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        cyclopentyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        n-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        cyclopentyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(isonipecotoyloxy)phenylalanine        ethyl ester    -   N-(α-toluenesulfonyl)-L-prolyl-L-4-(N-methylisonipecotoyloxy)phenylalanine        ethyl ester    -   N-(α-toluenesulfonyl)-L-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-3-(N,N-dimethylcarbamyloxy)phenylalanine        ethyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(1-tert-butylcarbonyloxy-4-phenylpiperidin-4-ylcarbonyloxy)phenylalanine        ethyl ester    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-[(1,1-dioxo)thiamorpholin-3-carbonyl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-[(1,1-dioxo)thiamorpholin-3-carbonyl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)sarcosyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)sarcosyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)sarcosyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylaminosulfonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylaminosulfonyloxy)phenylalanine    -   N-(1-methylimidazole-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-aminobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)sarcosyl-L-4-(morpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(morpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(α-t6luenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(piperazin-2-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(α-toluenesulfonyl)-L-(5,5-dimethyl)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        N-adamantyl amide    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanylglycine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylaminosulfonyloxy)phenylalanine        methyl ester    -   N-(toluene-4-sulfonyl)-L-(piperazin-2-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(4-benzyloxycarbonylpiperazin-2-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)sarcosyl-L-4-(isonipecotoyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L- [(1,1-dioxo)thiamorpholin-3        -carbonyl]-L-4-(morpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-[(1,1-dioxo)thiamorpholin-3-carbonyl]-L-4-(morpholin-4-ylcarbonyloxy)phenylalanine    -   N-(1-methylpyrazole-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)sarcosyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxo-5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(1-methylimidazole-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   2-(saccharin-2-yl)propionoyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxo-5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(pyridine-3-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-D-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-N-methylalanyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-nitrobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-[(1,1-dioxo)thiamorpholin-3-carbonyl]-L-4-(N,N-dimethylaminosulfonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)sarcosyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-N-methylalanyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(1,1-dioxothiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(isonipecotoyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(pyrrolidin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        neopentyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        neopentyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-tert-butyloxycarbonylpiperazin-1-ylcarbonyloxy)phenylalanine        ethyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(morpholin-4-ylcarbonyloxy)phenylalanine        ethyl ester    -   2-(saccharin-2-yl)propionoyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   2-(saccharin-2-yl)propionoyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)sarcosyl-L-4-(1,1-dioxothiomorphblin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)sarcosyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-N-methylalanyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)sarcosyl-L-4-(1,1-dioxothiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(morpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-N-methylalanyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(pyridine-3-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(pyrimidine-2-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-nitrobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-cyanobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylaminosulfonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxo)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxo)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(piperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(1-tert-butyloxycarbonylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(piperazin-1-ylcarbonyloxy)phenylalanine        ethyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-acetylpiperazin-1-ylcarbonyloxy)phenylalanine        ethyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-methanesulfonylpiperazin-1-ylcarbonyloxy)phenylalanine        ethyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(morpholin-4-ylcarbonyloxy)-3-nitrophenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(1-tert-butyloxycarbonylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-N-methyl-2-(tert-butyl)glycinyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(1,1-dioxothiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(1,1-dioxothiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-prolyl-L-4-(1,1-dioxothiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-prolyl-L-4-(morpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(morpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-trifluoromethoxybenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   3-[N-(toluene-4-sulfonyl)-N-methylamino]-1-[1-tert-butyloxycarbonyl-2-(N,N-dimethylcarbamyloxy)phenylethyl]azetidin-2-one    -   N-(4-fluorobenzenesulfonyl)-L-(1,1-dioxo-5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxo-5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(morpholin-4-ylcarbonyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(pyrimidine-2-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   3-[N-(toluene-4-sulfonyl)-N-methylamino]-1-[1-carboxy-2-(N,N-dimethylcarbamyloxy)phenylethyl]azetidin-2-one    -   N-(1-methylpyrazole-4-sulfonyl)-L-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(1,1-dioxo)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(isonipecotoyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(1,1-dioxothiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(pyrrolidin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(1,1-dioxo)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(2,5-dichlorothiophene-3-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-acetamidobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimetbylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-tert-butylbenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(pyridine-2-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(2-fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(3        -fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(2,4-difluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-acetamidobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-trifluoromethoxybenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-cyanobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(3,3-dimethyl)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(3,3-dimethyl)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        iso-propyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N-(1,4-dioxa-8-aza-spiro[4.5]decan-8-yl)carbonyloxy)phenylalanine        ethyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N-(1,4-dioxa-8-aza-spiro[4.5]decan-8-yl)carbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4′-acetylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4′-methanesulfonylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4′-phenylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(piperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   2-(saccharin-2-yl)propionyl-L-4-(4′-methylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4′-methanesulfonylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        (N ′-tert-butoxycarbonyl-2-amino-2-methylpropyl) ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4′-acetylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-hydroxypiperidin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N-(2-(morpholin-4′-yl)ethyl)carbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N-(1,4-dioxa-8-aza-spiro[4.5]decan-8-yl)carbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N-(2-hydroxyethyl)-N-methylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-4-(4′-(2-hydroxyethyl)piperazin-        1 -ylcarbonyloxy)-L-phenylalanine tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N-(2-fornyloxyethyl)-N-methylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N-(2′-hydroxyethyl)-N-methylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toulene-4-sulfonyl)-L-prolyl-L-4-(N-(methoxycarbonylmethyl)carbamyloxy)phenylalanine        tert-butyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-(4-N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4′-methoxypiperidin-1-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4′-methoxypiperidin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-4-oxoprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-trans-4-hydroxyprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(3-fluorobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(morpholino-sulfonyl)-L-prolyl-L-(4-N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(morpholino-sulfonyl)-L-prolyl-L-(4-N,N-dimethylcarbamyloxy)phenylalanine    -   N-(1-methylpyrazole-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(2-fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(2,4-difluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(pyridine-3-sulfonyl)-L-(5,5-dimethyl-thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3-fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(1-methylpyrazole-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-tert-butylbenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-(3,3-dimethyl)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(2,5-dichlorothiophene-3-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-methoxybenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-methoxybenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-(1-oxo-thiomorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(1-oxo-thiomorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(3,4-difluorobenzenesulfonyl)-L-prolyl-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3,4-difluorobenzenesulfonyl)-L-prolyl-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(3,4-difluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(3,4-difluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-(thiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        ethyl ester    -   N-(pyridine-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(pyridine-2-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(pyridine-2-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(pyridine-2-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(pyridine-2-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(2-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3,4-difluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3,5-difluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(2,4-difluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(4-chlorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3-chlorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(2-chlorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3,4-dichlorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3,5-dichlorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3        -chlorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(3,4-dichlorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-methoxybenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3-methoxybenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(2-methoxybenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3,4-dimethoxybenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(2,4-difluorobenzenesulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3,4-dichlorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(3-chlorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(3-chloro-4-fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(3,4-difluorobenzenesulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thioprolyl-L-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(3,4-difluorobenzenesulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(2,5-dichlorothiophene-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(8-quinolinesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(8-quinolinesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(8-quinolinesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isoproplyl ester    -   N-(8-quinolinesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-phenylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4′-(ethoxycarbonyl)piperidin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(pyridine-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(3-sulfonamido-4-chloro-benzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-(1-oxothiomorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(2,4-difluorobenzenefulfonyl)-L-(1-oxothiomorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        2,2-dimethylpropyl ester    -   N-(pyridine-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        2,2-dimethylpropyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        cyclopropylmethyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        methyl ester    -   N-(pyridine-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        ethyl ester    -   N-(pyridine-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        cyclopropylmethyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        2-methoxyphenyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        n-butyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        n-propyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        2,2-dimethylpropionyloxymethyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N-(4′-(2′-aminoethyl)morpholino)carbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-[4-(carboxy)piperidin-1-ylcarbonyloxy]phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-bis-(2-hydroxyethyl)carbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-[3-(hydroxymethyl)piperidin-1-ylcarbonyloxy]phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-trifluoromethanesulfonylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-(N-phenylurea)benzenesuilfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(2-trifluoroacetyl-1,2,3,4-tetrahydroisoquinolin-7-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(1-methylpyrazole-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(1-methylpyrazole-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(pyridine-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(pyridine-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N-methyl-N-(2-dimethylaminoethyl)carbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N-methyl-N-(2-dimethylaminoethyl)carbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thiapropyl-L-4-(N-methyl-N-(2-dimethylaminoethyl)carbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N-methyl-N-(2-dimethylaminoethyl)carbamyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3        -chloro-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(N,N-dimethycarbamyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(N,N-dimethylcarbamyloxy)]phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-methylpiperazin-1-ylcarbonyloxy)]phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-3-chloro-4-(N,N-dimethylcarbamyloxy)]phenylalanine        isopropyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-(2′-pyridyl)-piperazin-1-ylcarbonyloxy)]phenylalanine        isopropyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-(2′-pyridyl)-piperazin-1-ylcarbonyloxy)]phenylalanine        tert-butyl ester    -   N-(4-nitrobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(4-aminobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-phenylcarbamylpiperazin-1-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-phenylcarbamylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(1-n-butylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(pyridin-4-ylcarbonyl)piperazin-1-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-4-oxoprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-trans-4-hydroxyprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-cyanobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(4-aminobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-4-oxoprolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-[3-(hydroxymethyl)piperidin-1-ylcarbonyloxy]phenylalanine    -   N-(toluene-4-sulfonyl)-L-(4,4-difluoro)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-(4,4-difluoro)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-(4-benzoylpiperazin-1-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(1-methyl-1H-imidazole-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-4-(thiomorpholin-4-ylcarbonyloxy)prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(4-cyanobenzenesulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(4-amidinobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        methyl ester    -   N-(toluene-4-sulfonyl)-L-4-oxoprolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-4-hydroxyprolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-(4-benzoylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(4-amidinobenzenesulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        methyl ester    -   N-(3-fluorobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-[N-methyl-N-(2-(N′-methyl-N′-toluenesulfonyl-amino)ethyl)carbamyloxy]phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-[N-(2-(N′-phenylaminocarbonyloxy)ethyl)carbamyloxy)]phenylalanine        isopropyl ester    -   N-(4-fluorobenzenesulfonyl)-L-4-(trans-hydroxy)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(4-fluorobenzenesulfonyl)-L-4-(trans-hydroxy)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-amidinobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   piperazine-1,4-dicarboxylic acid        bis-{4-[(2S)-2-tert-butoxycarbonyl-2-((4R)-5,5-dimethyl-3-(toluene-4-sulfonyl)thiazolidine-4-carboxamido)ethyl]phenyl}        ester    -   piperazine-1,4-dicarboxylic acid        bis-{4-[(2S)-2-carboxy-2-((4R)-5,5-dimethyl-3-(toluene-4-sulfonyl)thiazolidine-4-carboxamido)ethyl]phenyl}        ester    -   N-(toluene-4-sulfonyl)-L-(pyrazin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(2-hydroxymethylpyrrolidin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(2-hydroxymethylpyrrolidin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(2-methoxycarbonylpyrrolidin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)]phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)]phenylalanine        tert-butyl ester    -   piperazine-1,4-dicarboxylic acid        bis-{4-[(2S)-2-isopropoxycarbonyl-2-((2R)-1-(toluene-4-sulfonyl)pyrrolidine-2-carboxamido)ethyl]phenyl}        ester    -   N-(toluene-4-sulfonyl)-L-(4-hydroxy)prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        2-(2-methoxyethoxy)ethyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyrimidyl)piperazin-1-ylcarbonyloxy)]phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3        -fluoro-4-(N,N-dimethylcarbamyloxy)phenylalanine isopropyl ester    -   N-(toluene-4-sulfonyl)-L-(1-methanesulfonylpyrazin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-[2-(1,1-dioxo-2,3-dihydro-3,3-dimethyl-1,2-benzisothiazol-2-yl)acetyl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-[2-(N-2,10-camphorsultamyl)acetyl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-[2-(N-2,10-camphorsultamyl)acetyl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-[2-(N-2,10-camphorsultamyl)acetyl]-L-3-chloro-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(4-bromobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-bromobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(4-hydroxy)prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyrimidyl)piperazin-1-ylcarbonyloxy)]phenylalanine    -   piperazine-1,4-dicarboxylic acid        bis-{4-[(2S)-2-tert-butoxycarbonyl-2-((2R)-1-(toluene-4-sulfonyl)pyrrolidine-2-carboxamido)ethyl]phenyl}        ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)]phenylalanine        isopropyl ester    -   N-(4-fluorobenzenesulfonyl)thiazolidinyl-2-carbonyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)thiazolidinyl-2-carbonyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(4-oxo)prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(4-oxo)prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)thiazolidinyl-2-carbonyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)]phenylalanine    -   N-(4-nitrobenzenesulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)]phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)thiazolidinyl-2-carbonyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)]phenylalanine        tert-butyl ester    -   N-(4-bromobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)]phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-(N-phenylthiocarbonyl)piperazin-1-ylcarbonyloxy)]phenylalanine        isopropyl ester    -   N-(4-fluorobenzenesulfonyl)th        iazolidinyl-2-carbonyl-L-4-(4-methylhomopiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   piperazine-1,4-dicarboxylic acid        bis-{4-[(2S)-2-carboxyl-2-((2R)-1-(toluene-4-sulfonyl)pyrrolidine-2-carboxamido)ethyl]phenyl}        ester    -   N-(toluene-4-sulfonyl)-L-4-(methanesulfonyloxy)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-aminocarbonylbenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-aminocarbonylbenzenesulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(4-amidinobenzenesulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(4-nitrobenzenesulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)]phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)]phenylalanine        ethyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)thiazolidinyl-2-carbonyl-L-4-(4-methylhomopiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(1-methylpyrazole-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(1-methylimidazole-4-sulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(1-methylimidazole-4-sulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylala        nine tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(4-fluorobenzenesulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(4-fluorobenzenesulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(1-methanesulfonylpyrazin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-4-(methanesulfonyloxy)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(methanesulfonyl)-N-benzylglycinyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-bromobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-trifluoromethoxybenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-trifluoromethoxybenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-trifluoromethoxybenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(4-hydroxy)prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine    -   N-(4-trifluoromethoxybenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine    -   N-(1-methylimidazole-4-sulfonyl)-L-prolyl-L-3-chloro-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(1-methylimidazole-4-sulfonyl)-L-prolyl-L-3-chloro-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(1-methylimidazole-4-sulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine    -   N-(1-methylimidazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine    -   N-(1-methylpyrazole-3-sulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine    -   N-(1-methylpyrazole-3-sulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(1-methylpyrazole-3-sulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(1-methylpyrazole-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(1-methylimidazole-4-sulfonyl)-L-prolyl-L-3-chloro-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(1-methylpyrazole-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        2-phenoxyethyl ester    -   N-(1-methylpyrazole-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine    -   N-(1-methylpyrazole-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        ethyl ester    -   N-(3-chloro-1,5-dimethylpyrazole-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-(5-trifluoromethyl-2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        and pharmaceutically acceptable salts thereof as well as any of        the ester compounds recited above wherein one ester is replaced        with another ester selected from the group consisting of methyl        ester, ethyl ester, n-propyl ester, isopropyl ester, n-butyl        ester, isobutyl ester, sec-butyl ester, tert-butyl ester and        neopentyl ester.

More preferred compounds within the scope of Formulae I and IA and IB(below) include by way of example:

-   -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        ethyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        ethyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        n-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        cyclopentyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        n-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        cyclopentyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(isonipecotoyloxy)phenylalanine        ethyl ester    -   N-(α-toluenesulfonyl)-L-prolyl-L-4-(N-methylisonipecotoyloxy)phenylalanine        ethyl ester    -   N-(α-toluenesulfonyl)-L-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-3-(N,N-dimethylcarbamyloxy)phenylalanine        ethyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(1-tert-butylcarbonyloxy-4-phenylpiperidin-4-ylcarbonyloxy)phenylalanine        ethyl ester    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-[(1,1-dioxo)thiamorpholin-3-carbonyl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-[(1,1-dioxo)thiamorpholin-3-carbonyl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)sarcosyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)sarcosyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)sarcosyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(1-methylimidazole-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-aminobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)sarcosyl-L-4-(morpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(morpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(α-toluenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(piperazin-2-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(α-toluenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(piperazin-2-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(4-benzyloxycarbonylpiperazin-2-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)sarcosyl-L-4-(isonipecotoyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-[(1,1-dioxo)thiamorpholin-3-carbonyl]-L-4-(morpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfotiyl)-L-[(1,1-dioxo)thiamorpholin-3-carbonyl]-L-4-(morpholin-4-ylcarbonyloxy)phenylalanine    -   N-(1-methylpyrazole-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)sarcosyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxo-5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(1-methylimidazole-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxo-5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(pyridine-3        -sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-D-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-N-methylalanyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-nitrobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)sarcosyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-N-methylalanyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(1,1-dioxothiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(isonipecotoyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(pyrrolidin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(morpholin-4-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        neopentyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        neopentyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-tert-butyloxycarbonylpiperazin-1-ylcarbonyloxy)phenylalanine        ethyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(morpholin-4-ylcarbonyloxy)phenylalanine        ethyl ester    -   N-(toluene-4-sulfonyl)sarcosyl-L-4-(1,1-dioxothiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)sarcosyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-N-methylalanyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)sarcosyl-L-4-(1,1-dioxothiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(morpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-N-methylalanyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(pyridine-3-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(pyrimidine-2-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-nitrobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-cyanobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxo)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxo)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(piperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(1-tert-butyloxycarbonylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(piperazin-1-ylcarbonyloxy)phenylalanine        ethyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-acetylpiperazin-1-ylcarbonyloxy)phenylalanine        ethyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-methanesulfonylpiperazin-1-ylcarbonyloxy)phenylalanine        ethyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(morpholin-4-ylcarbonyloxy)-3-nitrophenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(1-tert-butyloxycarbonylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-N-methyl-2-(tert-butyl)glycinyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(1,1-dioxothiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl        )-L-4-(1,1-dioxothiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-prolyl-L-4-(1,1-dioxothiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-prolyl-L-4-(morpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(morpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-trifluoromethoxybenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   3-[N-(toluene-4-sulfonyl)-N-methylamino]-1-[1-tert-butyloxycarbonyl-2-(N,N-dimethylcarbamyloxy)phenylethyl]azetidin-2-one    -   N-(4-fluorobenzenesulfonyl)-L-(1,1-dioxo-5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxo-5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(morpholin-4-ylcarbonyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(pyrimidine-2-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   3-[N-(toluene-4-sulfonyl)-N-methylamino]-1-[1-carboxy-2-(N,N-dimethylcarbamyloxy)phenylethyl]azetidin-2-one    -   N-(1-methylpyrazole-4-sulfonyl)-L-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(1,1-dioxo)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(isonipecotoyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(1,1-dioxothiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(pyrrolidin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(1,1-dioxo)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(2,5-dichlorothiophene-3-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-acetamidobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-tert-butylbenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(pyridine-2-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(2-fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(3-fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(2,4-difluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3        -carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine tert-butyl        ester    -   N-(4-acetamidobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-trifluoromethoxybenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-cyanobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(3,3-dimethyl)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(3,3-dimethyl)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        iso-propyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N-(1,4-dioxa-8-aza-spiro[4.5]decan-8-yl)carbonyloxy)phenylalanine        ethyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N-(1,4-dioxa-8-aza-spiro[4.5]decan-8-yl)carbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4′-acetylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4′-methanesulfonylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4′-phenylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(piperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4′-methanesulfonylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        (N,N-tert-butoxycarbonyl-2-amino-2-methylpropyl) ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4′-acetylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-hydroxypiperidin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N-(2-(morpholin-4′-yl)ethyl)carbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N-(1,4-dioxa-8-aza-spiro[4.5]decan-8-yl)carbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N-(2-hydroxyethyl)-N-methylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-4-(4′-(2-hydroxyethyl)piperazin-1-ylcarbonyloxy)-L-phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N-(2-formyloxyethyl)-N-methylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N-(2′-hydroxyethyl)-N-methylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toulene-4-sulfonyl)-L-prolyl-L-4-(N-(methoxycarbonylmethyl)carbamyloxy)phenylalanine        tert-butyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-(4-N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4′-methoxypiperidin-1-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4′-methoxypiperidin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-4-oxoprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-trans-4-hydroxyprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(3-fluorobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(morpholino-sulfonyl)-L-prolyl-L-(4-N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(morpholino-sulfonyl)-L-prolyl-L-(4-N,N-dimethylcarbamyloxy)phenylalanine    -   N-(1-methylpyrazole-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(2-fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(2,4-difluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(pyridine-3-sulfonyl)-L-(5,5-dimethyl-thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3-fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(1-methylpyrazole-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-tert-butylbenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-(3,3-dimethyl)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(2,5-dichlorothiophene-3-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-methoxybenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-methoxybenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-(1-oxo-thiomorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(1-oxo-thiomorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(3        ,4-difluorobenzenesulfonyl)-L-prolyl-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3,4-difluorobenzenesulfonyl)-L-prolyl-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(3,4-difluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(3,4-difluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-(thiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        ethyl ester    -   N-(pyridine-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(pyridine-2-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(pyridine-2-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(pyridine-2-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(pyridine-2-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(2-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3,4-difluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3,5-difluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(2,4-difluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(4-chlorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3-chlorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbarnyloxy)phenylalanine        isopropyl ester    -   N-(2-chlorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3,4-dichlorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3,5-dichlorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3-chlorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(3,4-dichlorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-methoxybenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3-methoxybenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(2-methoxybenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3,4-dimethoxybenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(2,4-difluorobenzenesulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(3,4-dichlorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(3-chlorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(3-chloro-4-fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(3,4-difluorobenzenesulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thioprolyl-L-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(3,4-difluorobenzenesulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(2,5-dichlorothiophene-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(8-quinolinesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(8-quinolinesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(8-quinolinesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isoproplyl ester    -   N-(8-quinolinesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-phenylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4′-(ethoxycarbonyl)piperidin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(pyridine-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(3-sulfonamido-4-chloro-benzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-(1-oxothiomorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(2,4-difluorobenzenefulfonyl)-L-(1-oxothiomorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        2,2-dimethylpropyl ester    -   N-(pyridine-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        2,2-dimethylpropyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        cyclopropylmethyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        methyl ester    -   N-(pyridine-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        ethyl ester    -   N-(pyridine-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbarniyloxy)phenylalanine        cyclopropylmethyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        2-methoxyphenyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        n-butyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        n-propyl ester    -   N-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        2,2-dimethylpropionyloxymethyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N-(4′-(2′-aminoethyl)morpholino)carbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-[4-(carboxy)piperidin-1-ylcarbonyloxy]phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-bis-(2-hydroxyethyl)carbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-[3-(hydroxymethyl)piperidin-1-ylcarbonyloxy]phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-trifluoromethanesulfonylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-(N-phenylurea)benzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(2-trifluoroacetyl-1,2,3,4-tetrahydroisoquinolin-7-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(1-methylpyrazole-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(1-methylpyrazole-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(pyridine-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(pyridine-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N-methyl-N-(2-dimethylaminoethyl)carbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N-methyl-N-(2-dimethylaminoethyl)carbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thiapropyl-L-4-(N-methyl-N-(2-dimethylaminoethyl)carbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N-methyl-N-(2-dimethylaminoethyl)carbamyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(N,N-dimethycarbamyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(N,N-dimethylcarbamyloxy)]phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-methylpiperazin-1-ylcarbonyloxy)]phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-3-chloro-4-(N,N-dimethylcarbamyloxy)]phenylalanine        isopropyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-(2′-pyridyl)-piperazin-1-ylcarbonyloxy)]phenylalanine        isopropyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-(2′-pyridyl)-piperazin-1-ylcarbonyloxy)]phenylalanine        tert-butyl ester    -   N-(4-nitrobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(4-aminobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-phenylcarbamylpiperazin-1-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-phenylcarbamylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(1-n-butylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(pyridin-4-ylcarbonyl)piperazin-1-ylcarbonyloxy)phenylalanine        isopropyl ester.    -   N-(toluene-4-sulfonyl)-L-4-oxoprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-trans-4-hydroxyprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-cyanobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(4-aminobenzenesulfonyl)-L-prolyl-L-4-(N,        N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-4-oxoprolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-[3-(hydroxymethyl)piperidin-1-ylcarbonyloxy]phenylalanine    -   N-(toluene-4-sulfonyl)-L-(4,4-difluoro)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-(4,4-difluoro)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-(4-benzoylpiperazin-1-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(1-methyl-1H-imidazole-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-4-(thiomorpholin-4-ylcarbonyloxy)prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(4-cyanob        enzenesulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(4-amidinobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        methyl ester    -   N-(toluene-4-sulfonyl)-L-4-oxoprolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-4-hydroxyprolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-(4-benzoylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(4-amidinobenzenesulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        methyl ester    -   N-(3-fluorobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-[N-methyl-N-(2-(N′-methyl-N′-toluenesulfonyl-amino)ethyl)carbamyloxy]phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-[N-(2-(N′-phenylaminocarbonyloxy)ethyl)carbamyloxy)]phenylalanine        isopropyl ester    -   N-(4-fluorobenzenesulfonyl)-L-4-(trans-hydroxy)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(4-fluorobenzenesulfonyl)-L-4-(trans-hydroxy)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-amidinobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(pyrazin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(2-hydroxymethylpyrrolidin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(2-hydroxymethylpyrrolidin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(2-methoxycarbonylpyrrolidin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3        -chloro-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)]phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)]phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(4-hydroxy)prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        2-(2-methoxyethoxy)ethyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyrimidyl)piperazin-1-ylcarbonyloxy)]phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-fluoro-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(toluene-4-sulfonyl)-L-(1-methanesulfonylpyrazin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-bromobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-bromobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(4-hydroxy)prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyrimidyl)piperazin-1-ylcarbonyloxy)]phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)]phenylalanine        isopropyl ester    -   N-(4-fluorobenzenesulfonyl)thiazolidinyl-2-carbonyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)thiazolidinyl-2-carbonyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(4-oxo)prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-(4-oxo)prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)thiazolidinyl-2-carbonyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)]phenylalanine    -   N-(4-nitrobenzenesulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)]phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)th        iazolidinyl-2-carbonyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)]phenylalanine        tert-butyl ester    -   N-(4-bromobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)]phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-(N-phenylthiocarbonyl)piperazin-1-ylcarbonyloxy)]phenylalanine        isopropyl ester    -   N-(4-fluorobenzenesulfonyl)thiazolidinyl-2-carbonyl-L-4-(4-methylhomopiperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-4-(methanesulfonyloxy)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-aminocarbonylbenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-aminocarbonylbenzenesulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(4-amidinobenzenesulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine    -   N-(4-nitrobenzenesulfonyl)-L-pro        lyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)]phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)]phenylalanine        ethyl ester    -   N-(4-fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)th        iazolidinyl-2-carbonyl-L-4-(4-methylhomopiperazin-1-ylcarbonyloxy)phenylalanine    -   N-(1-methylpyrazole-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(1-methylimidazole-4-sulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(1-methylimidazole-4-sulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(4-fluorobenzenesulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(4-fluorobenzenesulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(toluene-4-sulfonyl)-L-(        -methanesulfonylpyrazin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(toluene-4-sulfonyl)-L-4-(methanesulfonyloxy)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-bromobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-trifluoromethoxybenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-trifluoromethoxybenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester    -   N-(4-trifluoromethoxybenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(4-fluorobenzenesulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine    -   N-(4-fluorobenzenesulfonyl)-L-(4-hydroxy)prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine    -   N-(4-trifluoromethoxybenzenesulfonyl)-L-(5,5        -dimethyl)thiaprolyl-L-4-(4-(²-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine    -   N-(1-methylimidazole-4-sulfonyl)-L-prolyl-L-3-chloro-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(1-methylimidazole-4-sulfonyl)-L-prolyl-L-3-chloro-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester    -   N-(1-methylimidazole-4-sulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine    -   N-(1-methylimidazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine    -   N-(1-methylpyrazole-3-sulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine    -   N-(1-methylpyrazole-3-sulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(1-methylpyrazole-3-sulfonyl)-L-prolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(1-methylpyrazole-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        tert-butyl ester    -   N-(1-methylimidazole-4-sulfonyl)-L-prolyl-L-3-chloro-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        isopropyl ester    -   N-(1-methylpyrazole-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine-2-phenoxyethyl        ester    -   N-(1-methylpyrazole-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine    -   N-(1-methylpyrazole-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-(2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        ethyl ester    -   N-(3-chloro-1,5-dimethylpyrazole-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-(5-trifluoromethyl-2-pyridyl)piperazin-1-ylcarbonyloxy)phenylalanine        and pharmaceutically acceptable salts thereof.

Preferred compounds of Formulae I and IA above include those set forthin Table 1 below: TABLE 1

R¹ R² R³ R⁵ R⁶ p-CH₃-φ- R²/R³ = cyclicp-[(4-methylpiperazin-1-yl)C(O)O—]benzyl- —OCH₂CH₃ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl-—OCH₂CH₃ 3 carbon atoms (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-methylpiperazin-1-yl)C(O)O—]benzyl- —OCH(CH₃)₂ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-methylpiperazin-1-yl)C(O)O—]benzyl- —O-n-butyl 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-methylpiperazin-1-yl)C(O)O—]benzyl- —O-cyclopentyl 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-methylpiperazin-1-yl)C(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-methylpiperazin-1-yl)C(O)O—]benzyl- —OH 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl-—OCH(CH₃)₂ 3 carbon atoms (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —O-n-butyl 3 carbon atoms (L-pyrrolidinyl)p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —O-cyclopentyl 3 carbonatoms (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl-—OC(CH₃)₃ 3 carbon atoms (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl) p-CH₃-φ-R²/R³ = cyclic p-[(piperidin-4-yl)C(O)O—]benzyl- —OCH₂CH₃ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(1-methylpiperidin-4-yl)C(O)O—]benzyl- —OCH₂CH₃ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-methylpiperazin-1-yl)C(O)O—]benzyl- —OH 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclic m-[(CH₃)₂NC(O)O—]benzyl-—OCH₂CH₃ 3 carbon atoms (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(1-Boc-4-phenylpiperidin-4-yl)-C(O)O- —OCH₂CH₃ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl-—OCH(CH₃)₂ —CH₂S—C(CH₃)₂— (L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ-R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂CH₂—SO₂—CH₂— (L-1,1-dioxothiomorpholin-3-yl) p-CH₃-φ- R²/R³ = cyclic p-[(4-methylpiperazin-1-yl)C(O)O—]benzyl-—OC(CH₃)₃ —CH₂—S—C(CH₃)₂— (L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ-R²/R³ = cyclic p-[(4-methylpiperazin-1-yl)C(O)O—]benzyl- —OH—CH₂—S—C(CH₃)₂— (L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ- CH₃— Hp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ p-CH₃-φ- CH₃— Hp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ p-CH₃-φ- CH₃— Hp-[(CH₃)₂NC(O)O—]benzyl- —OH 1-methylimidazol-4-yl R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms (L-pyrrolidinyl)p-NH₂-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ 3 carbonatoms (L-pyrrolidinyl) p-CH₃-φ- CH₃— H p-[(morpholin-4-yl)C(O)O—]benzyl-—OC(CH₃)₃ p-CH₃-φ- R²/R³ = cyclic p-[(morpholin-4-yl)C(O)O—]benzyl-—OC(CH₃)₃ 3 carbon atoms (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂CH₂—NH—CH₂— (L-piperazinyl) p-CH₃-φ-R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—S—(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂CH₂—NH—CH₂— (L-piperazinyl)p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃—CH₂CH₂—(Cbz)NHCH₂— [L-4-N-(Cbz)-piperazinyl] p-CH₃-φ- CH₃— Hp-[(piperidin-1-yl)C(O)O—]benzyl- —OH p-CH₃-φ- R²/R³ = cyclicp-[(morpholin-4-yl)C(O)O—]benzyl- —OC(CH₃)₃ —CH₂CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) p-CH₃-φ- R²/R³ = cyclicp-[(morpholin-4-yl)C(O)O—]benzyl- —OH —CH₂CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) 1-methylpyrazol-4-yl R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms (L-pyrrolidinyl) p-F-φR²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ —CH₃ Hp-[(4-methylpiperazin-1-yl)C(O)O—]benzyl- —OC(CH₃)₃ p-CH₃-φ R²/R³ =cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—SO₂—C(CH₃)₂—(L-1,1-dioxo-5,5- dimethylthiazolidin-4-yl) 1-methylpyrazol-4-yl R²/R³ =cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl)p-CH₃-φ R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—SO₂—C(CH₃)₂—(L-1,1-dioxo-5,5- dimethylthiazolidin-4-yl) p-F-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl) 3-pyridylR²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclicp-[(4-methylpiperazin-1-yl)C(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms(D-pyrrolidinyl) p-CH₃-φ —CH₃ —CH₃p-[(4-methylpiperazin-1-yl)C(O)O—]benzyl- —OC(CH₃)₃ p-nitro-φ R²/R³ =cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ —CH₃ H p-[(thiomorpholin-4-yl)C(O)O—]benzyl-—OC(CH₃)₃ p-CH₃-φ —CH₃ —CH₃ p-[(4-methylpiperazin-1-yl)C(O)O—]benzyl-—OH p-CH₃-φ R²/R³ = cyclic p-[(thiomorpholin-4-ylsulfone)-C(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl) p-CH₃-φR²/R³ = cyclic p-[(thiomorpholin-4-yl)C(O)O—]benzyl- —OH 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclicp-[(piperidin-1-yl)C(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl)p-CH₃-φ R²/R³ = cyclic p-[(pyrrolidin-1-yl)C(O)O—]benzyl- —OH 3 carbonatoms (L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclicp-[(morpholin-4-yl)C(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl)p-CH₃-φ R²/R³ = cyclic p-[(4-methylpiperazin-1-yl)C(O)O—]benzyl-—OCH₂C(CH₃)₃ 3 carbon atoms (L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH₂C(CH₃)₃ 3 carbon atoms (L-pyrrolidinyl)p-CH₃-φ R²/R³ = cyclic p-[(1-Boc-piperazin-4-yl)C(O)O—]benzyl- —OCH₂CH₃3 carbon atoms (L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclicp-[(morpholin-4-yl)C(O)O—]benzyl- —OCH₂CH₃ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ —CH₃ H p-[(thiomorpholin-4-ylsulfone)-C(O)O—]benzyl- —OC(CH₃)₃ p-CH₃-φ —CH₃ Hp-[(thiomorpholin-4-yl)C(O)O—]benzyl- —OH p-CH₃-φ —CH₃ —CH₃p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ p-CH₃-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—CH₂—S—CH₂—(L-thiomorpholin-3-yl) p-CH₃-φ —CH₃ H p-[(thiomorpholin-4-ylsulfone)-C(O)O—]benzyl- —OH p-CH₃-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) p-CH₃-φ R²/R³ = cyclicp-[(morpholin-4-yl)C(O)O—]benzyl- —OC(CH₃)₃ —CH₂—CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) p-CH₃-φ —CH₃ —CH₃p-[(CH₃)₂NC(O)O—]benzyl- —OH p-F-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—CH₂—S—CH₂—(L-thiomorpholin-3-yl) p-F-φ R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl-—OC(CH₃)₃ —CH₂—CH₂—SO₂—CH₂— (L-1,1-dioxothiomorpholin- 3-yl)pyridin-3-yl R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ 3 carbonatoms (L-pyrrolidinyl) p-nitro-φ R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl-—OH 3 carbon atoms (L-pyrrolidinyl) p-N≡C-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms (L-pyrrolidinyl)p-CH₃-φ R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) p-CH₃-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—SO₂—CH₂—(L-1,1-dioxothiazolidin-4-yl) p-F₃C-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms (L-pyrrolidinyl)1-methylpyrazol-4-yl R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH 3carbon atoms (L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—SO₂—CH₂— (L-1,1-dioxothiazolidin-4-yl)p-F-φ R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—S—CH₂—(L-thiazolidin-4-yl) p-CH₃-φ R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl-2,4-dioxo- 3 carbon atoms tetrahydrofuran- (L-pyrrolidinyl)3-yl(3,4-enol) p-CH₃-φ R²/R³ = cyclic p-[(piperazin-4-yl)C(O)O—]benzyl-—OH 3 carbon atoms (L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclicp-[(1-Boc-piperazin-4-yl)C(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclicp-[(piperazin-4-yl)C(O)O—]benzyl- —OCH₂CH₃ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclicp-[(4-acetylpiperazin-1-yl)C(O)O—]benzyl- —OCH₂CH₃ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclicp-[(4-methanesulfonylpiperazin-1-yl)-C(O)O—] —OCH₂CH₃ 3 carbon atomsbenzyl- (L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclic3-nitro-4-[(morpholin-4-yl)-C(O)O—]benzyl- —OH 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclicp-[(1-Boc-piperazin-4-yl)C(O)O—]benzyl- —OH 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ —CH₃ —C(CH₃)₃p-[(4-methylpiperazin-1-yl)C(O)O—]benzyl- —OC(CH₃)₃ p-F-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-F-φ R²/R³ = cyclicp-[(1,1-dioxothiomorpholin-4-yl)- —OC(CH₃)₃ —CH₂—CH₂—SO₂—CH₂—C(O)O—]benzyl- (L-1,1-dioxothiomorpholin- 3-yl) p-CH₃-φ R²/R³ = cyclicp-[(1,1-dioxothiomorpholin-4-yl)- —OC(CH₃)₃ —CH₂—CH₂—SO₂—CH₂—C(O)O—]benzyl- (L-1,1-dioxothiomorpholin- 3-yl) p-F-φ R²/R³ = cyclicp-[(1,1-dioxothiomorpholin-4-yl)- —OC(CH₃)₃ 3 carbon atomsC(O)O—]benzyl- (L-pyrrolidinyl) p-F-φ R²/R³ = cyclicp-[(thiomorpholin-4- —OC(CH₃)₃ 3 carbon atoms yl)C(O)O—]benzyl-(L-pyrrolidinyl) p-F-φ R²/R³ = cyclic p-[(morpholin-4-yl)C(O)O—]benzyl-—OC(CH₃)₃ 3 carbon atoms (L-pyrrolidinyl) p-F-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-F-φ R²/R³ = cyclicp-[(morpholin-4-yl)C(O)O—]benzyl- —OC(CH₃)₃ —CH₂—CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) p-F₃CI-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms (L-pyrrolidinyl)p-CH₃-φ R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂—CH₂—CH₂—SO₂—CH₂— (L-1,1-dioxothiomorpholin- 3-yl) p-F-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—SO₂—C(CH₃)₂— (L-1,1-dioxo-5,5-dimethylthiazolidin-4-yl) p-CH₃-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—SO₂—C(CH₃)₂— (L-1,1-dioxo-5,5-dimethylthiazolidin-4-yl) p-CH₃-φ R²/R³ = cyclicp-[(morpholin-4-yl)C(O)O—]benzyl- —OH —CH₂—CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) p-F-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—SO₂—C(CH₃)₂— (L-1,1-dioxo-5,5-dimethylthiazolidin-4-yl) pyrimidin-2-yl R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl) p-CH₃-φR²/R³ = cyclic p-[(4-methylpiperazin-1-yl)C(O)O—]benzyl- —OC(CH₃)₃—CH₂—CH₂—SO₂—CH₂— (L-1,1-dioxothiomorpholin- 3-yl) p-F-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂SO₂—CH₂—(L-1,1-dioxothiazolidin-4-yl) 2,5-dichlorothien-3-yl R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms (L-pyrrolidinyl)p-CH₃C(O)NH-φ R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ 3 carbonatoms (L-pyrrolidinyl) p-C(CH₃)₃-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) pyridin-2-yl R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl) o-F-φ R²/R³= cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) m-F-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) 2,4-difluoro-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) p-CH₃C(O)NH-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl) p-C(F)₃O-φR²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms(L-pyrrolidinyl) p-F-φ R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl-—OCH(CH₃)₂ —CH₂—S—C(CH₃)₂— (L-5,5-dimethylthiazolidin-4- yl) p-N≡C-φR²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms(L-pyrrolidinyl) morpholin-4-yl R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl-—OC(CH₃)₃ 3 carbon atoms (L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—CH₂—C(CH₃)₂— (L-4,4-dimethylpyrrolidinyl) p-CH₃-φ R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂—CH₂—CH₂—C(CH₃)₂— (L-4,4-dimethyl pyrrolidinyl) 1-methylpyrazol-4-ylR²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl-—OCH(CH₃)₂ —CH₂—S—C(CH₃)₂— (L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ-R²/R³ = cyclic p-[(thiomorpholin-4-yl)C(O)O—]benzyl- —OC(CH₃)₃ 3 carbonatoms (L-pyrrolidinyl) 1-methylimidazol-4-yl R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl) p-CH₃-φ-R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) 1-methylpyrazol-4-yl R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-CH₃C(O)NH-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms (L-pyrrolidinyl)p-(CH₃)₃C-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃—CH₂—CH₂—SO₂—CH₂— (L-1,1-dioxothiomorpholin- 3-yl) 1-methylpyrazol-4-ylR²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) pyridin-3-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl) p-N≡C-φ-R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms(L-pyrrolidinyl) p-F-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl-—OCH(CH₃)₂ —CH₂—S—C(CH₃)₂— (L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ-R²/R³ = cyclic p-[(1,4-dioxa-8-azaspiro[4.5]decan- —OCH₂CH₃ 3 carbonatoms 8-yl)-C(O)O—]benzyl- (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(1,4-dioxa-8-azaspiro[4.5]decan- —OH 3 carbon atoms8-yl)-C(O)O—]benzyl (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-acetylpiperazin-1-yl)C(O)O—]benzyl- —OH 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-methanesulfonylpiperazin-1-yl)-C(O)O—] —OH 3 carbon atoms benzyl-(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-p-piperazin-1-yl)C(O)(O)—]benzyl- —OH 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(piperazin-1-yl)C(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl-—OC(CH₃)₃ —CH₂CH₂—NH—CH₂— (L-piperazinyl) p-F₃CO-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl) p-CH₃-φ-R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—CH₂—C(CH₃)₂—(4,4-dimethyl pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—CH₂—C(CH₃)₂— (4,4-dimethylpyrrolidinyl) p-CH₃C(O)NH-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH3 carbon atoms (L-pyrrolidinyl) o-F-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) morpholin-4-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms (L-pyrrolidinyl)m-F-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃—CH₂—CH₂—SO₂—CH₂— (L-1,1-dioxothiomorpholin- 3-yl) 2,4-difluoro-φ- R²/R³= cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) morpholin-4-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl)1-methylpyrazol-4-yl R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃—CH₂—CH₂—SO₂—CH₂— (L-1,1-dioxothiomorpholin- 3-yl) o-F-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) 2,4-difluoro-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—CH₂—S—CH₂— (L-thiomorpholin-3-yl)pyridin-3-yl- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂—CH₂—S—C(CH₃)₂— (L-5,5-dimethylthiazolidin-4- yl) m-F-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) pyridin-2-yl R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ 3 carbon atoms (L-pyrrolidinyl)1-methylpyrazol-4-yl R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH(L-1,1-dioxothiomorpholin- 3-yl) p-CH₃-φ- R²/R³ = cyclicp-[(4-methanesulfonylpiperazin-1-yl)-C(O)O—] —OC(CH₃)₃ 3 carbon atomsbenzyl- (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-p-piperazin-1-yl)C(O)(O)-]benzyl- —OC(CH₃)₃ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —O— 3carbon atoms CH₂C(CH₃)₂— (L-pyrrolidinyl) NHC(O)OC(CH₃)₃ p-CH₃-φ- R²/R³= cyclic p-[(CH₃)₂NC(O)O—]benzyl- —O—CH₂CH₂— 3 carbon atoms(morpholin-4-yl) (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-acetylpiperazin-1-yl)C(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NCH₂CH₂N(CH₃)C(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NCH₂CH₂N(CH₃)C(O)O—]benzyl- —OC(CH₃)₃ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NCH₂CH₂N(CH₃)C(O)O—]benzyl- —OH —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NCH₂CH₂N(CH₃)C(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl)p-CH₃-φ- R²/R³ = cyclic p-[(4-hydroxypiperidin-1-yl)C(O)O—]benzyl-—OC(CH₃)₃ 3 carbon atoms (L-pyrrolidinyl) p-(CH₃)₃C-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) p-CH₃-φ- R²/R³ = cyclicp-[(thiomorpholin-4-yl)C(O)O—]benzyl- —OC(CH₃)₃ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 2,5-dichlorothien-3-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl) p-CH₃O-φ-R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH—CH₂—CH₂—C(CH₃)₂— (4,4-dimethyl pyrrolidinyl) p-F-φ- R²/R³ = cyclic3-chloro-4-[(4-methylpiperazin-1-yl)C(O)O —OC(CH₃)₃ —CH₂—S—C(CH₃)₂—benzyl- (L-5,5-dimethylthiazolidin-4- yl) p-F-φ- R²/R³ = cyclic3-chloro-4-[(CH₃)₂NC(O)Obenzyl- —OH —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-F-φ- R²/R³ = cyclic3-chloro-4-[(thiomorpholin-4-yl)C(O)Obenzyl- —OC(CH₃)₃ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ- R²/R³ = cyclicp-[(morpholin-4-yl)-CH₂CH₂NHC(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(1,4-dioxa-8-azaspiro[4.5]decan- —OC(CH₃)₃ 3 carbon atoms8-yl)-C(O)O—]benzyl- (L-pyrrolidinyl) p-F-φ- R²/R³ = cyclic3-chloro-4-[(CH₃)₂NC(O)Obenzyl- —OC(CH₃)₃ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ- R²/R³ = cyclicp-[(thiomorpholin-4-yl)C(O)O—]benzyl- —OH —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) pyridin-2-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl) p-CH₃-φ-R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—CH₂—(O)—CH₂—(L-1-oxothiomorpholin-4-yl) 4-Cl-3-(NH₂—SO₂φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ 3 carbon atoms (L-pyrrolidinyl)p-F-φ- R²/R³ = cyclic 3-chloro-4-[(thiomorpholin-4-yl)C(O)Obenzyl-—OCH(CH₃)₂ —CH₂—S—C(CH₃)₂— (L-5,5-dimethylthiazolidin-4- yl) p-F-φ-R²/R³ = cyclic 3-chloro-4-[(CH₃)₂NC(O)Obenzyl- —OCH(CH₃)₂—CH₂—S—C(CH₃)₂— (L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ- R²/R³ =cyclic p-[HOCH₂CH₂N(CH₃)C(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms(L-pyrrolidinyl) p-F-φ- R²/R³ = cyclicp-[(2-(hydroxymethyl)pyrrolidin-1-yl)-C(O)O —OC(CH₃)₃ —CH₂—S—C(CH₃)₂—benzyl- (L-5,5-dimethylthiazolidin-4- yl) p-F-φ- R²/R³ = cyclicp-[(2-(hydroxymethyl)pyrrolidin-1-yl)-C(O)O —OH —CH₂—S—C(CH₃)₂— benzyl-(L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ- R²/R³ = cyclicp-[(2-(CH₃OC(O)—)pyrrolidin- —OC(CH₃)₃ 3 carbon atoms1-yl)-C(O)Obenzyl- (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-(HC(O)O—)piperidin-1-yl)-C(O)Obenzyl- —OH 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-(hydroxypiperidin-1-yl)-C(O)Obenzyl- —OCH(CH₃)₂ 3 carbon atoms(L-pyrrolidinyl) p-F-φ- R²/R³ = cyclic3-chloro-4-[(thiomorpholin-4-yl)C(O)Obenzyl- —OH —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ- R²/R³ = cyclicp-[(4-(CH₃CH₂OC(O)—)piperidin-1-yl)C(O)O —OC(CH₃)₃ 3 carbon atomsbenzyl- (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-(HOCH₂CH₂—)piperazin-1-yl)-C(O)O— —OC(CH₃)₃ 3 carbon atoms(L-pyrrolidinyl) ]benzyl- p-F-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)-C(O)O— —OH —CH₂—S—C(CH₃)₂— benzyl-(L-5,5-dimethylthiazolidin-4- ]benzyl yl) p-F-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)-C(O)O—] —OC(CH₃)₃ —CH₂—S—C(CH₃)₂—benzyl- (L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ- R²/R³ = cyclicp-[HC(O)OCH₂CH₂N(CH₃)C(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl)p-CH₃-φ- R²/R³ = cyclic p-[HOCH₂CH₂N(CH₃)C(O)O—]benzyl- —OCH(CH₃)₂ 3carbon atoms (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[CH₃OC(O)CH₂NHC(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms (L-pyrrolidinyl)quinolin-8-yl- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ 3carbon atoms (L-pyrrolidinyl) 3,4-difluoro-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ 3 carbon atoms (L-pyrrolidinyl)p-CH₃-φ- R²/R³ = cyclic p-[(thiomorpholin-4-yl)-C(O)O—]benzyl-—OCH(CH₃)₂ 3 carbon atoms (L-pyrrolidinyl) p-CH₃O-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl) p-CH₃-φ-R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—CH₂—(O)—CH₂—(L-1-oxothiomorpholin-4-yl) 3,4-difluoro-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) p-H₂N-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ 3 carbon atoms (L-pyrrolidinyl)3,4-difluoro-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH—CH₂CH₂—SO₂—CH₂— (L-1,1-dioxothiomorpholin- 3-yl) 3,4-difluoro-φ- R²/R³= cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl)quinolin-8-yl- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbonatoms (L-pyrrolidinyl) 1-methylpyrazol-4-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—CH₂—(O)—CH₂—(L-1-oxothiomorpholin-4-yl) 1-n-butylpyrazol-4-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 1-methylpyrazol-3-yl R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 2-(CF₃C(O)1,2,3,4- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ tetrahydro-isoquinolin- 3 carbonatoms 7-yl (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-(φNHC(O)piperazin-1-yl)-C(O)O—] —OH 3 carbon atoms benzyl-(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-methoxypiperidin-1-yl)C(O)O—]benzyl- —OH 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-(pyridin-4-ylC(O))piperazin-1-yl)-C(O)O— —OCH(CH₃)₂ 3 carbon atomsbenzyl- (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—C(O)—CH₂— (L-4-oxopyrrolidinyl)p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃—CH₂—CH(OH)CH₂— (L-4-hydroxopyrrolidinyl) m-F-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms (L-pyrrolidinyl)p-CH₃-φ- R²/R³ = cyclic p-[(4-methoxypiperidin-1-yl)C(O)O—]benzyl-—OCH(CH₃)₂ 3 carbon atoms (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-(pNHC(O)—)piperazin-1- —OCH(CH₃)₂ 3 carbon atomsyl)-C(O)O—]benzyl- (L-pyrrolidinyl) p-(φNHC(O)NH)φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ 3 carbon atoms (L-pyrrolidinyl)p-F-φ- R²/R³ = cyclic 3-chloro-4-[(4-methylpiperidin-1-yl)-C(O)O—OCH(CH₃)₂ —CH₂—S—C(CH₃)₂— benzyl- (L-5,5-dimethylthiazolidin-4- benzyl-yl) p-CH₃-φ- R²/R³ = cyclic 3-chloro-4-[(CH₃)₂NC(O)Obenzyl- —OCH(CH₃)₂—CH₂—S—C(CH₃)₂— (L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ- R²/R³ =cyclic p-[(4-(CH₃SO₂—)piperazin-1-yl)-C(O)O]benzyl- —OC(CH₃)₃ 3 carbonatoms (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(morpholin-4-yl)CH₂CH₂NHC(O)O—]benzyl- —OH 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-(HO(O)—)piperidin-1-yl)C(O)O—]benzyl- —OH 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclic p-[(HOCH₂CH₂)₂NC(O)O—]benzyl-—OCH(CH₃)₂ 3 carbon atoms (L-pyrrolidinyl) p-O₂N-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ 3 carbon atoms (L-pyrrolidinyl)p-CH₃-φ- R²/R³ = cyclic p-[(4-(HOCH₂—)piperidin-1-yl)- —OCH(CH₃)₂ 3carbon atoms C(O)O—]benzyl- (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclic3-chloro-4-[(CH₃)₂NC(O)O)benzyl- —OCH(CH₃)₂ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl-—OCH(CH₃)₂ —CH₂—CH₂—S—CH₂— (L-thiomorpholin-3-yl) 1-methylpyrazol-3-ylR²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) m-F-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) o-F-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 3,4-difluoro-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 3,5-difluoro-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 2,4-difluoro-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-NH₂-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl) p-N≡C-φ-R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH—CH₂CH(OH)CH₂— (L-4-hydroxypyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂C(O)CH₂— (L-4-oxypyrrolidinyl)pyridin-2-yl- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂—CH₂—S—C(CH₃)₂— (L-5,5-dimethylthiazolidin-4- yl) p-Cl-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) m-Cl-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) o-Cl-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 3,4-dichloro-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 3,5-dichloro-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 1-methylpyrazol-4-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH₂CH₃ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) pyridin-3-yl R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 1-methylpyrazol-4-yl R²/R³ = cyclicp-[(thiomorpholin-4-yl)—C(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) quinolin-8-yl R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) m-Cl-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) pyridin-2-yl R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 3,4-dichloro-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) 2,5-dichlorothien-3-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-CH₃O-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) m-CH₃O-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) o-CH₃O-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 3,4-dimethoxy-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 2,4-difluoro-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—CH₂—S—CH₂—(L-thiomorpholin-3-yl) 3,4-dichloro-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂CH₂—SO₂—CH₂— (L-1,1-dioxothiomorpholin-3-yl) m-Cl-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH—CH₂CH₂—SO₂—CH₂— (L-1,1-dioxothiomorpholin- 3-yl) 2,4-difluoro-φ- R²/R³= cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—CH₂—S(O)—CH₂—(L-1-oxothiomorpholin-4-yl) p-CH₃-φ- R²/R³ = cyclicp-[(4-methylpiperzin-1-yl)C(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—C(O)—CH₂—(L-4-oxopyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(thiomorpholin-4yl)-C(O)O—]benzyl- —OC(CH₃)₃ —CH₂CH(OH)CH₂—(L-4-hydroxypyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(3-(HOCH₂—)piperidin-1- —OH 3 carbon atoms yl)-C(O)O—]benzyl-(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl-—OCH(CH₃)₂ —CH₂—CF₂—CH₂— (L-4,4-difluoro-pyrrolidinyl) p-CH₃-φ- R²/R³ =cyclic p-[(CH₃)₂NC(O)O—]benzyl- O(CH₂CH₂O)₂CH₃ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(thiomorpholin-4-yl)-C(O)O—]benzyl- —OC(CH₃)₃ —CH₂CH(—O—C(O)thiomorpholin-4-yl- CH₂— (L-4-(thiomorpholin-4-yl)C(O)O-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl-—OH —CH₂—CF₂—CH₂— (L-4,4-difluoro-pyrolidinyl) p-F-φ- R²/R³ = cyclicp-[(4-(pyrimidin-2-yl)piperazin-1-yl)-C(O)O—] —OC(CH₃)₃ —CH₂—S—C(CH₃)₂—benzyl- (L-5,5-dimethylthiazolidin-4- ]benzyl yl) p-CH₃-φ- R²/R³ =cyclic p-[(4-(pC(O)—)piperazin-1-yl)-C(O)O—]benzyl- —OCH(CH₃)₂ 3 carbonatoms (L-pyrrolidinyl) p-F-φ- R²/R³ = cyclic3-fluoro-4-[(CH₃)₂NC(O)Obenzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-F-φ- R²/R³ = cyclic3-chloro-4-[(4-(pyridin-2-yl)piperazin-1- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—yl)C(O)Obenzyl- (L-5,5-dimethylthiazolidin-4- yl) p-F-φ- R²/R³ = cyclic3-chloro-4-[(4-(pyridin-2-yl)piperazin-1- —OC(CH₃)₃ —CH₂—S—C(CH₃)₂—yl)C(O)Obenzyl- (L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ- R²/R³ =cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂CH₂NSO₂CH₃)—CH₂—(L-4-methanesulfonyl- piperazinyl) 1-mehylimidazol-4- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ yl- 3 carbon atoms (L-pyrrolidinyl)p-Br-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃—CH₂—S—C(CH₃)₂— (L-5,5-dimethylthiazolidin-4- yl) p-Br-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-NH₂C(═N)-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH₃ 3 carbon atoms (L-pyrrolidinyl) p-N≡C-φ-R²/R³ = cyclic p-[(thiomorpholin-4-yl)-C(O)O—]benzyl- —OCH(CH₃)₂ 3carbon atoms (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(thiomorpholin-4-yl)-C(O)O—]benzyl- —OH —CH₂CH(—O—C(O)thiomorpholin-4-yl)- CH₂— (L-4-(thiomorpholin-4-yl)C(O)O-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(thiomorpholin-4-yl)-C(O)O—]benzyl- —OC(CH₃)₃ —CH₂—C(O)—CH₂—(L-4-oxopyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(thiomorpholin-4-yl)-C(O)O—]benzyl- —OH —CH₂—C(O)—CH₂—(L-4-oxopyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-methylpiperazin-1-yl)-C(O)O—]benzyl- —OH —CH₂—C(O)—CH₂—(L-4-oxopyrrolidinyl) p-F-φ- R²/R³ = cyclicp-[(4-(pyrimidin-2-yl)piperazin-1-yl)-C(O)O—] —OH —CH₂—S—C(CH₃)₂—benzyl- (L-5,5-dimethylthiazolidin-4- yl) quinolin-8-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) pyridin-4-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-F-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—yl)C(O)O—]benzyl- (L-5,5-dimethylthiazolidin-4- yl) m-F-φ- R²/R³ =cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms (L-pyrrolidinyl)p-F-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—CH₂—S—(thiazolidin-2-yl) p-F-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O-Jbenzyl-—OC(CH₃)₃ —CH₂—CH₂—S— (thiazolidin-2-yl) p-CH₃-φ- R²/R³ = cyclicp-[(thiomorpholin-4-yl)C(O)O—]benzyl- —OH —CH₂—C(O)—CH₂—(L-4-oxopyrrolidinyl) p-NH₂—C(≡N)-φ- R²/R³ = cyclicp-[(thiomorpholin-4-yl)C(O)O—]benzyl- —OCH₃ 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-methylpiperazin-1-yl)-C(O)O—]benzyl- —OH —CH₂—C(O)—CH₂—(L-4-oxopyrrolidinyl) p-F-φ- R²/R³ = cyclicp-[(4-pyridin-2-yl)piperazin-1- —OH —CH₂—CH₂—S— yl)-C(O)O—]benzyl-(thiazolidin-2-yl) p-NO₂-φ- R²/R³ = cyclicp-[(4-pyridin-2-yl)piperazin-1- —OC(CH₃)₃ 3 carbon atomsyl)-C(O)O—]benzyl- (L-pyrrolidinyl) p-F-φ- R²/R³ = cyclicp-[(4-pyridin-2-yl)piperazin-1- —OC(CH₃)₃ —CH₂—CH₂—S— yl)-C(O)O—]benzyl-(thiazolidin-2-yl) p-Br-φ- R²/R³ = cyclicp-[(4-pyridin-2-yl)piperazin-1- —OH —CH₂—S—C(CH₃)₂— yl)-C(O)O—]benzyl-(L-5,5-dimethylthiazolidin-4- yl) p-CH₃-φ- R²/R³ = cyclicp-[(4-(qC(O)-)piperazin-1- —OH 3 carbon atoms yl)-C(O)O—]benzyl-(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclic p-[(4-(φNHC(S)—)piperazin-1-—OCH(CH₃)₂ 3 carbon atoms yl)-C(O)O—]benzyl- (L-pyrrolidinyl) p-F-φ-R²/R³ = cyclic p-[(4-CH₃-homopiperazin-1-yl)C(O)O—]benzyl- —OC(CH₃)₃—CH₂—CH₂—S— (thiazolidin-2-yl) p-CH₃-φ- R²/R³ = cyclic p-[p-CH₃-φ-—OCH(CH₃)₂ 3 carbon atoms SO₂N(CH₃)CH₂CH₂N(CH₃)—C(O)O— (L-pyrrolidinyl)benzyl- p-CH₃-φ- R²/R³ = cyclic p-[φNHC(O)O—CH₂CH₂NHC(O)O—]benzyl-—OCH(CH₃)₂ 3 carbon atoms (L-pyrrolidinyl) 3-Cl-4-F-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂CH₂—SO₂—CH₂—(L-1,1-dioxothiomorpholin- 3-yl) 1-methylpyrazol-4-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂CH₂—S—CH₂— (L-thiomorpholin-3-yl)p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃—CH₂CH(—OSO₂CH₃)—CH₂— (L-4-methanesulfoxy- pyrrolidinyl) p-H₂NC(O)-φ-R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH 3 carbon atoms(L-pyrrolidinyl) p-H₂N—C(═N)-φ R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl-—OH 3 carbon atoms (L-pyrrolidinyl) p-H₂NC(O)-φ- R²/R³ = cyclicp-[(thiomorpholin-4-yl)C(O)O—]benzyl- —OH 3 carbon atoms(L-pyrrolidinyl) p-H₂N—C(═N)-φ R²/R³ = cyclicp-[(thiomorpholin-4-yl)C(O)O—]benzyl- —OH 3 carbon atoms(L-pyrrolidinyl) p-NO₂-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)-C(O)O—] —OH 3 carbon atoms benzyl-(L-pyrrolidinyl) p-F-φ- R²/R³ = cyclic3-chloro-4-[(4-(pyridin-2-yl)piperazin-1- —OCH₂CH₃ —CH₂—S—C(CH₃)₂—yl)C(O)Obenzyl- (L-5,5-dimethylthiazolidin-4- yl) p-F-φ- R²/R³ = cyclic3-chloro-4-[(4-(pyridin-2-yl)piperazin-1- —OH —CH₂—S—C(CH₃)₂—yl)C(O)Obenzyl- (L-5,5-dimethylthiazolidin-4- yl) p-F-φ- R²/R³ = cyclicp-[(4-CH₃-homopiperazin-1-yl)C(O)Obenzyl- —OH —CH₂—CH₂—S—(thiazolidin-2-yl) 1-methylpyrazol-4-yl- R²/R³ = cyclic3-chloro-4-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 1-methylimidazol-4- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1- —OCH(CH₃)₂ 3 carbon atomsyl)C(O)O—]benzyl- yl- (L-pyrrolidinyl) 1-methylimidazol-4- R²/R³ =cyclic p-[(4-(pyridin-2-yl)piperazin-1- —OC(CH₃)₃ 3 carbon atomsyl)C(O)O—]benzyl- yl- (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1- —OH 3 carbon atoms yl)C(O)O—]benzyl-(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1- —OC(CH₃)₃ 3 carbon atomsyl)C(O)O—]benzyl- (L-pyrrolidinyl) p-F-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1- —OCH(CH₃)₂ 3 carbon atomsyl)C(O)O—]benzyl- (L-pyrrolidinyl) p-F-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1- —OCH(CH₃)₂ 3 carbon atomsyl)C(O)O—]benzyl- (L-pyrrolidinyl) p-F-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1- —OCH(CH₃)₂ 3 carbon atomsyl)C(O)O—]benzyl- (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂CH₂N(—SO₂—CH₃)CH₂— (4-methanesulfonyl-piperazin-2-yl) p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH—CH₂CH(—OSO₂—CH₃)CH₂— (L-4-methanesulfoxy- pyrrolidinyl) 3,4-difluoro-φ-R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—CH₂—S—CH₂—(L-thiomorpholin-3-yl) pyridin-3-yl R²/R³ = cyclicp-[(thiomorpholin-4-yl)C(O)O—]benzyl- —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 3,4-difluoro-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—CH₂—S—CH₂— (L-thiomorpholin-3-yl)p-F-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OCH(CH₃)₂ —CH₂CH(OH)CH₂—(L-4-hydroxypyrrolidinyl) p-Br-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1- —OC(CH₃)₃ —CH₂—S—C(CH₃)₂—yl)C(O)O—]benzyl- (L-5,5-dimethylthiazolidin-4- yl) p-CF₃O-φ- R²/R³ =cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-CF₃O-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) p-CF₃O-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1- —OC(CH₃)₃ —CH₂—S—C(CH₃)₂—yl)C(O)O—]benzyl- (L-5,5-dimethylthiazolidin-4- yl) p-F-φ R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1- —OH 3 carbon atoms yl)C(O)O—]benzyl-(L-pyrrolidinyl) p-F-φ- R²/R³ = cyclic p-[(4-(pyridin-2-yl)piperazin-1-—OH —CH₂CH(OH)CH₂— yl)C(O)O—]benzyl- (L-4-hydroxypyrrolidinyl) p-CF₃O-φ-R²/R³ = cyclic p-[(4-(pyridin-2-yl)piperazin-1- —OH —CH₂—S—C(CH₃)₂—yl)C(O)O—]benzyl- (L-5,5-dimethylthiazolidin-4- yl) 1-methylimidazol-4-R²/R³ = cyclic 3-chloro-4-[(CH₃)₂NC(O)O)-]benzyl- —OH 3 carbon atoms yl-(L-pyrrolidinyl) 1-methylimidazol-4- R²/R³ = cyclic3-chloro-4-[(CH₃)₂NC(O)O)-]benzyl- —OCH(CH₃)₂ 3 carbon atoms yl-(L-pyrrolidinyl) 1-methylimidazol-4- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1- —OH 3 carbon atoms yl)C(O)O—]benzyl-yl- (L-pyrrolidinyl) 1-methylimidazol-4- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1- —OH —CH₂—S—C(CH₃)₂— yl)C(O)O—]benzyl-yl- (L-5,5-dimethylthiazolidin-4- yl) 1-methylpyrazol-4-yl- R²/R³ =cyclic p-[(4-(pyridin-2-yl)piperazin-1- —OH 3 carbon atomsyl)C(O)O—]benzyl- (L-pyrrolidinyl) 1-methylpyrazol-4-yl- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1- —OCH(CH₃)₂ 3 carbon atomsyl)C(O)O—]benzyl- (L-pyrrolidinyl) 1-methylpyrazol-4-yl- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1- —OC(CH₃)₃ 3 carbon atomsyl)C(O)O—]benzyl- (L-pyrrolidinyl) 1-methylpyrazol-4-yl- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1- —OC(CH₃)₃ —CH₂—S—C(CH₃)₂—yl)C(O)O—]benzyl- (L-5,5-dimethylthiazolidin-4- yl) 1-methylimidazol-4-R²/R³ = cyclic 3-chloro-4-[(4-(pyridin-2-yl)piperazin-1- —OCH(CH₃)₂ 3carbon atoms yl)C(O)O]benzyl- yl- (L-pyrrolidinyl) 1-methylpyrazol-4-yl-R²/R³ = cyclic p-[(CH₃)₂NC(O)O—]benzyl- —OCH₂CH₂Oφ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 1-methylpyrazol-4-yl R²/R³ = cyclic3-chloro-4-[(4-(pyridin-2-yl)piperazin-1- —OH(L-5,5-dimethylthiazolidin-4- yl)C(O)Obenzyl- yl) 1-methylpyrazol-4-yl-R²/R³ = cyclic 3-chloro-4-[(4-(pyridin-2-yl)piperazin-1- —OCH₂CH₃—CH₂—S—C(CH₃)₂— yl)C(O)Obenzyl- (L-5,5-dimethylthiazolidin-4- yl)1,5-dimethyl-3- R²/R³ = cyclic p-[4-[5-CF₃-pyridin-2-yl)piperazin- —OHchloropyrazol-4-yl- —CH₂—S—C(CH₃)₂— 1 yl)-C(O)O—]benzyl-(L-5,5-dimethylthiazolidin-4- yl) p-F-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂CH(OH)CH₂—(L-4-hydroxypyrrolidinyl) pyridin-4-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OH —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 1-methylpyrazol-4-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH₂C(CH₃)₃ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 1-methylpyrazol-4-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH₂O- —CH₂—S—C(CH₃)₂— C(O)C(C(CH₃)₃(L-5,5-dimethylthiazolidin-4- yl) pyridin-3-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH₂C(CH₃)₃ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 1-methylpyrazol-4-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- 2-CH₃O-φ-O— —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 1-methylpyrazol-4-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- OCH₂cyclo- —CH₂—S—C(CH₃)₂— propyl(L-5,5-dimethylthiazolidin-4- yl) 1-methylpyrazol-4-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH₂CH₂CH₃ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 1-methylpyrazol-4-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH₂CH₂CH₂CH₃ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) 1-methylpyrazol-4-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —O—CH₃ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) pyridin-3-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OC(CH₃)₃ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) pyridin-3-yl- R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH₂CH₃ —CH₂—S—C(CH₃)₂—(L-5,5-dimethylthiazolidin-4- yl) pyridin-3-yl R²/R³ = cyclicp-[(CH₃)₂NC(O)O—]benzyl- —OCH₂cyclo- —CH₂—S—C(CH₃)₂— propyl(L-5,5-dimethylthiazolidin-4- yl)

In a preferred embodiment of the compounds of Formulae I and IA, thecompounds are defined by Formula IB below:

wherein:

-   -   Ar¹ is selected from the group consisting of aryl, substituted        aryl, heteroaryl, and substituted heteroaryl;    -   Ar² is selected from the group consisting of aryl, substituted        aryl, heteroaryl and substituted heteroaryl;

R¹² is selected from the group consisting of alkyl, substituted alkyl,cycloalkyl, and substituted cycloalkyl or R¹² and R¹³ together with thenitrogen atom bound to R¹² and the carbon atom bound to R¹³ form aheterocyclic or substituted heterocyclic group;

R¹³ is selected from the group consisting of hydrogen, alkyl andsubstituted alkyl, or R¹² and R¹³ together with the nitrogen atom boundto R¹² and the carbon atom bound to R¹³ form a heterocyclic orsubstituted heterocyclic group;

-   -   R¹⁴ is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, and        substituted aryl;    -   R¹⁵ is selected from the group consisting of alkyl, and        substituted alkyl, or R¹⁵ and R¹⁶ together with the nitrogen        atom to which they are bound form a heterocyclic or substituted        heterocyclic group;    -   R¹⁶ is selected from the group consisting of alkyl and        substituted alkyl or R¹⁵ and R¹⁶ together with the nitrogen atom        to which they are bound form a heterocyclic or substituted        heterocyclic group; and    -   is selected from the group consisting of —O—, —NR¹⁰⁰—, and —CH₂—        wherein R¹⁰⁰ is hydrogen or alkyl;    -   and pharmaceutically acceptable salts thereof.

Preferably, in the compounds of Formula IB above, R¹² is alkyl,substituted alkyl, or R¹² and R¹³ together with the nitrogen atom boundto R¹² and the carbon atom bound to R¹³ form a heterocyclic orsubstituted heterocyclic group. Preferably, in the compounds of FormulaIB above, R¹⁴ is hydrogen or alkyl.

Preferably, in the compounds of Formula IB above, Ar¹ is selected fromthe group consisting of phenyl, 4-methylphenyl, 4-t-butylphenyl,2,4,6-trimethylphenyl, 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl,2,4-difluorophenyl, 3,4-difluorophenyl, 3,5-difluorophenyl,2-chlorophenyl, 3-chlorophenyl, 4-chlorophenyl, 3,4-dichlorophenyl,3,5-dichlorophenyl, 3-chloro-4-fluorophenyl, 4-bromophenyl,2-methoxyphenyl, 3-methoxyphenyl, 4-methoxyphenyl, 3,4-dimethoxyphenyl,4-t-butoxyphenyl, 4-(3′-dimethylamino-n-propoxy)-phenyl,2-carboxyphenyl, 2-(methoxycarbonyl)phenyl, 4-(H₂NC(O)-)phenyl,4-(H₂NC(S)—)phenyl, 4-cyanophenyl, 4-trifluoromethylphenyl,4-trifluoromethoxyphenyl, 3,5-di-(trifluoromethyl)phenyl, 4-nitrophenyl,4-aminophenyl, 4-(CH₃C(O)NH-)phenyl, 4-(PhNHC(O)NH—)phenyl,4-amidinophenyl, 4-methylamidinophenyl, 4-[CH₃SC(═NH)-]phenyl,4-chloro-3-[H₂NS(O)₂-]phenyl, 1-naphthyl, 2-naphthyl, pyridin-2-yl,pyridin-3-yl, pyridine-4-yl, pyrimidin-2-yl, quinolin-8-yl,2-(trifluoroacetyl)-1,2,3,4-tetrahydroisoquinolin-7-yl, 2-thienyl,5-chloro-2-thienyl, 2,5-dichloro-4-thienyl, 1-N-methylimidazol-4-yl,1-N-methylpyrazol-3-yl, 1-N-methylpyrazol-4-yl, 1-N-butylpyrazol-4-yl,1-N-methyl-3-methyl-5-chloropyrazol-4-yl,1-N-methyl-5-methyl-3-chloropyrazol-4-yl, 2-thiazolyl and5-methyl-1,3,4-thiadiazol-2-yl.

Preferably, in the compounds of Formula IB above, R¹² and R¹³ togetherwith the nitrogen atom bound to R¹² and the carbon atom bound to R¹³form a heterocyclic or substituted heterocyclic of the formula:

-   -   wherein    -   X is selected from the group consisting of —S—, —SO—, —SO₂, and        optionally substituted —CH₂—;    -   m is an integer of 0 to 12;    -   n is an integer of 0 to 2; and    -   R′ is selected from the group consisting of alkyl, substituted        alkyl, and amino.

Preferably, m is 1, X is —S— or —CH₂—, R′ is alkyl or substituted alkyl.

Even more preferably, R¹² and R¹³ together with the nitrogen atom boundto R¹² and the carbon atom bound to R¹³ form a heterocyclic orsubstituted heterocyclic selected from the group consisting ofazetidinyl, thiazolidinyl, piperidinyl, piperazinyl, thiomorpholinyl,pyrrolidinyl, 4-hydroxypyrrolidinyl, 4-oxopyrrolidinyl,4-fluoropyrrolidinyl, 4,4-difluoropyrrolidinyl,4-(thiomorpholin-4-ylC(O)O—)pyrrolidinyl, 4-[CH₃S(O)₂O-]pyrrolidinyl,3-phenylpyrrolidinyl, 3-thiophenylpyrrolidinyl, 4-aminopyrrolidinyl,3-methoxypyrrolidinyl, 4,4-dimethylpyrrolidinyl, 4-N-Cbz-piperazinyl,4-[CH₃S(O)₂-]piperazinyl, thiazolidin-3-yl,5,5-dimethyl-thiazolidin-3-yl, 5,5-dimethylthiazolindin-4-yl,1,1-dioxo-thiazolidinyl, 1,1-dioxo-5,5-dimethylthiazolidin-2-yl and1,1-dioxothiomorpholinyl.

Preferably, in the compounds of Formula IB, Ar² is selected from thegroup consisting of phenyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, and4-pyrid-2-onyl.

Preferably, in Formula IB, Y is —O—, and when Y is —O—, the moiety—OC(O)NR¹⁵R¹⁶ is preferably selected from the group consisting of(CH₃)₂NC(O)O-, (piperidin-1-yl)C(O)O—, (4-hydroxypiperidin-1-yl)C(O)O—,(4-formyloxypiperidin-1-yl)C(O)O—,(4-ethoxycarbonylpiperidin-1-yl)C(O)O—,(4-carboxylpiperidin-1-yl)C(O)O—, (3-hydroxymethylpiperidin-1-yl)C(O)O—,(4-hydroxymethylpiperidin-1-yl)C(O)O—, (4-piperidon-1-yl ethyleneketal)C(O)O—, (piperazin-1-yl)-C(O)O—, (1-Boc-piperazin-4-yl)-C(O)O—,(4-methylpiperazin-1-yl)C(O)O—, (4-methylhomopiperazin-1-yl)C(O)O—,(4-(2-hydroxyethyl)piperazin-1-yl)C(O)O—,(4-phenylpiperazin-1-yl)C(O)O—, (4-(pyridin-2-yl)piperazin-1-yl)C(O)O—,(4-(4-trifluoromethylpyridin-2-yl)piperazin-1-yl)C(O)O—,(4-(pyrimidin-2-yl)piperazin-1-yl)C(O)O—,(4-acetylpiperazin-1-yl)C(O)O—, (4-(phenylC(O)-)piperazin-1-yl)C(O)O—,(4-(pyridin-4′-ylC(O)-)piperazin-1-yl)C(O)O,(4-(phenylNHC(O)-)piperazin-1-yl)C(O)O—,(4-(phenylNHC(S)-)piperazin-1-yl)C(O)O—,(4-methanesulfonylpiperazin-1-yl-C(O)O—,(4-trifluoromethanesulfonylpiperazin-1-yl-C(O)O—,(morpholin-4-yl)C(O)O—, (thiomorpholin-4-yl)C(O)O—, (thiomorpholin-4′-ylsulfone)-C(O)O-(pyrrolidin-1-yl)C(O)O—, (2-methylpyrrolidin-1-yl)C(O)O—,(2-(methoxycarbonyl)pyrrolidin-1-yl)C(O)O-,(2-(hydroxymethyl)pyrrolidin-1-yl)C(O)O—,(2-(N,N-dimethylamino)ethyl)(CH₃)NC(O)O—,(2-(N-methyl-N-toluene-4-sulfonylamino)ethyl)(CH₃)N—C(O)O—,(2-(morpholin-4-yl)ethyl)(CH₃)NC(O)O—, (2-(hydroxy)ethyl)(CH₃)NC(O)O—,bis(2-(hydroxy)ethyl)NC(O)O—, (2-(formyloxy)ethyl)(CH₃)NC(O)O—,(CH₃C(O)CH₂)HNC(O)O—, and 2-[(phenylNHC(O)O-)ethyl-]HNC(O)O—.

Preferably, the compound is the compound of Formula M below:

In a preferred embodiment, the compounds are defined by Formula IC below

wherein R^(x) is hydroxy or C₁₋₅ alkoxy and pharmaceutically acceptablesalts thereof. Preferably, the compound isN-[N-(3-pyridinesulfonyl)-L-3,3-dimethyl-4-thiaprolyl]-O-[I-methylpiperazin-4-ylcarbonyl]-L-tyrosineisopropyl ester (the compound of Formula N).

In another aspect, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compoundsdefined by Formula II below. These compounds have a binding affinity toVLA-4 as expressed by an IC₅₀ of about 15 μM or less (measured asdescribed in Example A below):

wherein:

-   -   R²¹ is selected from the group consisting of alkyl, substituted        alkyl, aryl, substituted aryl, cycloalkyl, substituted        cycloalkyl, heterocyclic, substituted heterocylic, heteroaryl        and substituted heteroaryl;    -   R²² is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, cycloalkyl, substituted cycloalkyl,        cycloalkenyl, substituted cycloalkenyl, heterocyclic,        substituted heterocyclic, aryl, substituted aryl, heteroaryl,        substituted heteroaryl, and R²¹ and R²² together with the        nitrogen atom bound to R²² and the SO₂ group bound to R²¹ can        form a heterocyclic or a substituted heterocyclic group;    -   R²³ is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl, substituted heteroaryl,        heterocyclic, substituted heterocyclic and where R²² and R²³        together with the nitrogen atom bound to R²² and the carbon atom        bound to R²³ can form a saturated heterocyclic group or a        saturated substituted heterocyclic group with the proviso that        when monosubstituted, the substituent on said saturated        substituted heterocyclic group is not carboxyl;    -   Q is —C(X)NR⁷— wherein R⁷ is selected from the group consisting        of hydrogen and alkyl;    -   X is selected from the group consisting of oxygen and sulfur;        and    -   R²⁵ is —CH₂Ar²²—R^(25′) where Ar²² is aryl or heteroaryl and R²⁵        is selected from the group consisting of aryl, heteroaryl,        substituted aryl, substituted heteroaryl, heterocyclic,        substituted heterocyclic, aryloxy, substituted aryloxy,        aralkoxy, substituted aralkoxy, heteroaryloxy, substituted        heteroaryloxy, heterocyclic-O—, substituted heterocyclic-O—,        heteroaralkoxy, and substituted heteroaralkoxy;    -   and pharmaceutically acceptable salts thereof.

In another embodiment, the compounds of this invention can also beprovided as prodrugs which convert (e.g., hydrolyze, metabolize, etc.)in vivo to a compound of Formula II above. In a preferred example ofsuch an embodiment, the carboxylic acid in the compound of Formula II ismodified into a group which, in vivo, will convert to the carboxylicacid (including salts thereof). In a particularly preferred embodiment,such prodrugs are represented by compounds of Formula IIA:

where

-   -   R²¹ is selected from the group consisting of alkyl, substituted        alkyl, aryl, substituted aryl, cycloalkyl, substituted        cycloalkyl, heterocyclic, substituted heterocylic, heteroaryl        and substituted heteroaryl;    -   R²² is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, cycloalkyl, substituted cycloalkyl,        cycloalkenyl, substituted cycloalkenyl, heterocyclic,        substituted heterocyclic, aryl, substituted aryl, heteroaryl,        and substituted heteroaryl, and R²¹ and R²² together with the        nitrogen atom bound to R²² and the SO₂ group bound to R²¹ can        form a heterocyclic or a substituted heterocyclic group;    -   R²³ is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl,        substituted aryl, heteroaryl, substituted heteroaryl,        heterocyclic, and substituted heterocyclic, and R²² and R²³        together with the nitrogen atom bound to R²² and the carbon atom        bound to R²³ can form a saturated heterocyclic group or a        saturated substituted heterocyclic group with the proviso that        when monosubstituted, the substituent on said saturated        substituted heterocyclic group is not carboxyl;    -   R²⁵ is —CH₂Ar²²—R^(25′) where Ar²² is aryl or heteroaryl and        R^(25′) is selected from the group consisting of aryl,        heteroaryl, substituted aryl, substituted heteroaryl,        heterocyclic, substituted heterocyclic, aryloxy, substituted        aryloxy, aralkoxy, substituted aralkoxy, heteroaryloxy,        substituted heteroaryloxy, , heterocyclic-O—, substituted        heterocyclic-O—, heteroaralkoxy, and substituted heteroaralkoxy;    -   R²⁶ is selected from the group consisting of        2,4-dioxo-tetrahydrofuran-3-yl (3,4-enol), amino, alkoxy,        substituted alkoxy, cycloalkoxy, substituted cyloalkoxy,        —O—(N-succinimidyl), —NH-adamantyl, —O-cholest-5-en-3-O-yl,        -NHOY where Y is hydrogen, alkyl, substituted alkyl, aryl, and        substituted aryl, —NH(CH₂)_(p)COOY where p is an integer of from        1 to 8 and Y is as defined above, —OCH₂NR²⁹R³⁰ where R²⁹ is        selected from the group consisting of —C(O)-aryl and        —C(O)-substituted aryl and R³⁰ is selected from the group        consisting of hydrogen and —CH₂COOR³¹ where R³¹ is alkyl, and        —NHSO₂Z′ where Z′ is alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, aryl, substituted aryl, heteroaryl,        substituted heteroaryl, heterocyclic or substituted        heterocyclic;    -   Q is —C(X)NR⁷— wherein R⁷ is selected from the group consisting        of hydrogen and alkyl; and    -   X is selected from the group consisting of oxygen and sulfur;    -   and pharmaceutically acceptable salts thereof.

Further description of the compounds of the above Formulae II and IIAand procedures and reaction conditions for preparing these compounds aredescribed in U.S. Ser. Nos. 09/127,346 (filed Jul. 31, 1998), 09/688,820(Continuation, filed Oct. 17, 2000 and issued as U.S. Pat. No.6,583,139) and 10/382,988 (Continuation, filed Mar. 7, 2003), all ofwhich are herein incorporated by reference in their entirety.

Preferably, in the compounds of Formulae II and IIA above, R²¹ isselected from the group consisting of alkyl, substituted alkyl, aryl,substituted aryl, heterocyclic, substituted heterocylic, heteroaryl andsubstituted heteroaryl. More preferably R²¹ is selected from the groupconsisting of aryl, substituted aryl, heteroaryl and substitutedheteroaryl.

Even more preferably, in the compounds of Formulae II and IIA above, R²¹s selected from the group consisting of 4-methylphenyl, 4-chlorophenyl,1-naphthyl, 2-naphthyl, 4-methoxyphenyl, phenyl, 2,4,6-trimethylphenyl,2-(methoxycarbonyl)phenyl, 2-carboxyphenyl, 3,5-dichlorophenyl,4-trifluoromethylphenyl, 3,4-dichlorophenyl, 3,4-dimethoxyphenyl,4-(CH₃C(O)NH—)phenyl, 4-trifluoromethoxyphenyl, 4-cyanophenyl,3,5-di-(trifluoromethyl)phenyl, 4-t-butylphenyl, 4-t-butoxyphenyl,4-nitrophenyl, 2-thienyl, 1-N-methyl-3-methyl-5-chloropyrazol-4-yl,1-N-methylimidazol-4-yl, 4-bromophenyl, 4-amidinophenyl,4-methylamidinophenyl, 4-[CH₃SC(═NH)]phenyl, 5-chloro-2-thienyl,2,5-dichloro-4-thienyl, 1-N-methy-1-4-pyrazolyl, 2-thiazolyl,5-methyl-1,3,4-thiadiazol-2-yl, 4-[H₂NC(S)]phenyl, 4-aminophenyl,4-fluorophenyl, 2-fluorophenyl, 3-fluorophenyl, 3,5-difluorophenyl,pyridin-3-yl, pyrimidin-2-yl, 4-(3′-dimethylamino-n-propoxy)-phenyl, and1-methylpyrazol-4-yl.

Preferably, R²², in the compounds of Formulae II and IIA above, ishydrogen, methyl, phenyl, benzyl, —(CH₂)₂-2-thienyl, and —(CH₂)₂-φ.

In another preferred embodiment, R²² and R²³, in the compounds ofFormulae II and IIA above, and R³² and R³³, in the compounds of FormulaIIB, together with the nitrogen atom bound to R²² or R³² and the carbonatom bound to R²³ or R³³ form a saturated heterocyclic group or asaturated substituted heterocyclic group with the proviso that whenmonosubstituted, the substituent on said saturated substitutedheterocyclic group is not carboxyl.

Q, in the compounds of Formulae II and IIA above, is preferably —C(O)NH—or —C(S)NH—.

In the compounds of Formulae II and IIA, R²⁵ is preferably selected fromthe group consisting of all possible isomers arising by substitutionwith the following groups: 4-(2-carboxyphenoxy)benzyl,4-(benzyloxy)benzyl, 4-[(1-methylpiperidin-4-yl)-O-]benzyl,4-(imidazolid-2-one-1-yl)benzyl, and4-(3-formylimidazolid-2-one-1-yl)benzyl.

In the compounds of Formula IIA, R²⁶ is preferably2,4-dioxo-tetrahydrofuran-3-yl (3,4-enol), methoxy, ethoxy, iso-propoxy,n-butoxy, t-butoxy, cyclopentoxy, neo-pentoxy,2-α-iso-propyl-4-β-methylcyclohexoxy,2-β-isopropyl-4-β-methylcyclohexoxy, —NH₂, benzyloxy, —NHCH₂COOH,—NHCH₂CH₂COOH, —NH-adamantyl, —NHCH₂CH₂COOCH₂CH₃, —NHSO₂-p-CH₃-φ, —NHOR⁸where R⁸ is hydrogen, methyl, iso-propyl or benzyl, O-(N-succinimidyl),—O-cholest-5-en-3-β-yl, —OCH₂—OC(O)C(CH₃)₃, —O(CH₂)_(z)NHC(O)W where zis 1 or 2 and W is selected from the group consisting of pyrid-3-yl,N-methylpyridyl, and N-methyl-1,4-dihydro-pyrid-3-yl, —NR″C(O)—R′ whereR′ is aryl, heteroaryl or heterocyclic and R″ is hydrogen or—CH₂C(O)OCH₂CH₃.

Preferred compounds within the scope of Formulae II and IIA aboveinclude by way of example the following:

-   -   N-(Toluene-4-sulfonyl)-L-prolyl-4-(α-methylbenzyloxy)-L-phenylalanine    -   N-(Toluene-4-sulfonyl)-L-prolyl-4-(2-carboxyphenoxy)-L-phenylalanine    -   N-(Toluene-4-sulfonyl)-L-prolyl-O-(benzyl)-L-tyrosine        and pharmaceutically acceptable salts thereof.

Preferred compounds of Formulae II and IIA above include those set forthin Table 2 below. TABLE 2

Q = —C(O)NR⁷— R²¹ R²² R²³ R²⁵ R²⁶ wherein R⁷ is: p-CH₃-φ- R²²/R²³ =cyclic p-[O-(o-carboxyphenyl)]-benzyl- —OH H 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²²/R²³ = cyclic p-benzyloxybenzyl- —OH H 3carbon atoms (L-pyrrolidinyl) p-CH₃-φ- R²²/R²³ = cyclicp-[(1-methylpiperidin-4-yl)-O-]benzyl- —OCH₂CH₃ H 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²²/R²³ = cyclicp-[(imidazolid-2-one-1-yl)benzyl- —OC(CH₃)₃ H 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²²/R²³ = cyclicp-[3-formylimidazolid-2-one-1-yl)benzyl- —OH H 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²²/R²³ = cyclicp-[(imidazolid-2-one-1-yl)benzyl- —OH H 3 carbon atoms (L-pyrrolidinyl)p-CH₃-φ- R²²/R²³ = cyclic p-[1-H-2-oxo-3-methyl tetrahydro —Ot-Bu H 3carbon atoms pyrimidin-1-yl]benzyl- (L-pyrrolidinyl) p-CH₃-φ- R²²/R²³ =cyclic p-[1-H-2-oxo-3-methyl tetrahydro —OH H 3 carbon atomspyrimidin-1-yl]benzyl- (L-pyrrolidinyl) p-CH₃-φ- R²²/R²³ = cyclic4-[2-methoxy phenyl]-benzyl- —Ot-Bu H 3 carbon atoms (L-pyrrolidinyl)p-CH₃-φ- R²²/R²³ = cyclic 4-[2-methoxy phenyl]-benzyl- —OH H 3 carbonatoms (L-pyrrolidinyl) p-CH₃-φ- R²²/R²³ = cyclicp-[2,4,5-trioxo-3-(3-chlorophenyl)- —OBz H 3 carbon atomstetrahydroimidazol-1-yl]-benzyl- (L-pyrrolidinyl)

In a preferred embodiment of the compounds of Formulae II and IIA, thecompounds are defined by Formula IIB below:

wherein:

-   -   Ar³¹ is selected from the group consisting of aryl, substituted        aryl, heteroaryl, and substituted heteroaryl;    -   R³² is selected from the group consisting of alkyl, substituted        alkyl, cycloalkyl, and substituted cycloalkyl or R³² and R³³        together with the nitrogen atom bound to R³² and the carbon atom        bound to R³³ form a heterocyclic or substituted heterocyclic        group;    -   R³³ is selected from the group consisting of hydrogen, alkyl,        and substituted alkyl, or R³² and R³³ together with the nitrogen        atom bound to R³² and the carbon atom bound to R³³ form a        heterocyclic or substituted heterocyclic group;    -   R³⁴ is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, and        substituted aryl; and    -   R³⁷ is aryl, heteroaryl, substituted aryl, substituted        heteroaryl, heterocyclic, substituted heterocyclic, aryloxy,        substituted aryloxy, aralkoxy, substituted aralkoxy,        heteroaryloxy, substituted heteroaryloxy;    -   and pharmaceutically acceptable salts thereof.

Preferably, in the compounds of Formula IIB above, R³² is alkyl,substituted alkyl, or R³² and R³³ together with the nitrogen atom boundto R³² and the carbon atom bound to R³³ form a heterocyclic orsubstituted heterocyclic group and R³⁴ is hydrogen or alkyl.

Preferably, in the compounds of Formula IIB above, R³⁷ is aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, orsubstituted heterocyclic. In a preferred embodiment, R³⁷ is substitutedaryl wherein the aryl is substituted with one to three substituentsindependently selected from the group consisting alkyl and alkoxy. In apreferred embodiment, R³⁷ is substituted heteroaryl wherein theheteroaryl is substituted with one to three substituents independentlyselected from the group consisting alkyl, alkoxy, and oxo. In anotherpreferred embodiment R³⁷ is substituted aryl or heteroaryl wherein arylor heteroaryl is 2,6-di-substituted. In yet another preferred embodimentR³⁷ is 2,6-di-substituted aryl wherein the substituents areindependently selected from the group consisting of alkyl. and alkoxy.In yet another preferred embodiment R³⁷ is 2,6-di-substituted heteroarylwherein the substituents are independently selected from the groupconsisting of alkyl, oxo, and alkoxy. In another preferred embodiment,R³⁷ is selected from the group consisting of 2,6-dialkoxyaryl,2,6-dialkoxyheteroaryl, 2-alkyl-6-alkoxyaryl,2-alkyl-6-alkoxyheteroaryl, 2-oxo-6-alkoxyheteroaryl,2-oxo-6-alkylheteroaryl, and optionally substitutedimidazolidin-2,4-dion-3-yl.

Preferably in the compounds of Formula IIB above, Ar³¹ is selected fromthe group consisting of 4-methylphenyl, 4-chlorophenyl, 1-naphthyl,2-naphthyl, 4-methoxyphenyl, phenyl, 2,4,6-trimethylphenyl,2-(methoxycarbonyl)phenyl, 2-carboxyphenyl, 3,5-dichlorophenyl,4-trifluoromethylphenyl, 3,4-dichlorophenyl, 3,4-dimethoxyphenyl,4-(CH₃C(O)NH—)phenyl, 4-trifluoromethoxyphenyl, 4-cyanophenyl,3,5-di-(trifluoromethyl)phenyl, 4-t-butylphenyl, 4-t-butoxyphenyl,4-nitrophenyl, 2-thienyl, 1-N-methyl-3-methyl-5-chloropyrazol-4-yl,1-N-methylimidazol-4-yl, 4-bromophenyl, 4-amidinophenyl,4-methylamidinophenyl, 4-[CH₃SC(═NH)]phenyl, 5-chloro-2-thienyl,2,5-dichloro-4-thienyl, 1-N-methyl-4-pyrazolyl, 2-thiazolyl,5-methyl-1,3,4-thiadiazol-2-yl, 4-[H₂NC(S)]phenyl, 4-aminophenyl,4-fluorophenyl, 2-fluorophenyl, 3-fluorophenyl, 3,5-difluorophenyl,pyridin-3-yl, pyrimidin-2-yl, 4-(3′-dimethylamino-n-propoxy)-phenyl, and1-methylpyrazol-4-yl.

When describing the compounds, compositions and methods of thisinvention, the following terms have the following meanings, unlessotherwise indicated.

Compound Preparation for Compounds of Formulae I and II

The compounds of Formulae I and II can be prepared from readilyavailable starting materials using the following general methods andprocedures. It will be appreciated that where typical or preferredprocess conditions (i.e., reaction temperatures, times, mole ratios ofreactants, solvents, pressures, etc.) are given, other processconditions can also be used unless otherwise stated. Optimum reactionconditions may vary with the particular reactants or solvent used, butsuch conditions can be determined by one skilled in the art by routineoptimization procedures.

Additionally, as will be apparent to those skilled in the art,conventional protecting groups may be necessary to prevent certainfunctional groups from undergoing undesired reactions. Suitableprotecting groups for various functional groups as well as suitableconditions for protecting and deprotecting particular functional groupsare well known in the art. For example, numerous protecting groups aredescribed in T. W. Greene and G. M. Wuts, Protecting Groups in OrganicSynthesis, Second Edition, Wiley, N.Y., 1991, and references citedtherein.

Furthermore, the compounds of this invention will typically contain oneor more chiral centers. Accordingly, if desired, such compounds can beprepared or isolated as pure stereoisomers, i.e., as individualenantiomers or diastereomers, or as stereoisomer-enriched mixtures. Allsuch stereoisomers.(and enriched mixtures) are included within the scopeof this invention, unless otherwise indicated. Pure stereoisomers (orenriched mixtures) may be prepared using, for example, optically activestarting materials or stereoselective reagents well-known in the art.Alternatively, racemic mixtures of such compounds can be separatedusing, for example, chiral column chromatography, chiral resolvingagents and the like.

According to the following compound preparation, R¹, R², R³, R⁵, R⁶, andR⁷ are as defined herein for Formulae I, IA, II, and IIA. In addition,according to the following compound preparation, R¹ is equivalent to:

-   -   Ar¹ as herein defined for Formula IB,    -   R²¹ as herein defined for Formulae II and IIA, and    -   Ar²¹ as herein defined for Formula IIB;        R² is equivalent to:    -   R¹² as herein defined for Formula IB,    -   R²² as herein defined for Formulae II and IIA, and    -   R³² as herein defined for Formula IIB;        R ³is equivalent to:    -   R¹³ as herein defined for Formula IB,    -   R²³ as herein defined for Formulae II and IIA, and    -   R³³ as herein defined for Formula IIB;        R⁵is equivalent to:    -   R²⁵ as herein defined for Formulae II and IIA; and        R⁶is equivalent to:    -   OH for Formulae I and II,    -   OR¹⁴ as herein defined for Formula IB,    -   R²⁶ as herein defined for Formula IIA, and    -   OR³⁴as herein defined for Formula IIB.

In a preferred method of synthesis, the compounds of Formulae I, IA, II,and IIA, wherein Q is —C(O)NR⁷—, and compounds of Formulae IB, IC, andIIB are prepared by first coupling an amino acid of Formula A:

with a sulfonyl chloride of Formula B:

to provide an N-sulfonyl amino acid of Formula C:

This reaction is typically conducted by reacting the amino acid ofFormula A with at least one equivalent, preferably about 1.1 to about 2equivalents, of sulfonyl chloride B in an inert diluent such asdichloromethane and the like. Generally, the reaction is conducted at atemperature ranging from about −70° C. to about 40° C. for about 1 toabout 24 hours. Preferably, this reaction is conducted in the presenceof a suitable base to scavenge the acid generated during the reaction.Suitable bases include, by way of example, tertiary amines, such astriethylamine, diisopropylethylamine, N-methylmorpholine and the like.Alternatively, the reaction can be conducted under Schotten-Baumann-typeconditions using aqueous alkali, such as sodium hydroxide and the like,as the base. Upon completion of the reaction, the resulting N-sulfonylamino acid C is recovered by conventional methods includingneutralization, extraction, precipitation, chromatography, filtration,and the like.

The amino acids of Formula A employed in the above reaction are eitherknown compounds or compounds that can be prepared from known compoundsby conventional synthetic procedures. Examples of suitable amino acidsfor use in this reaction include, but are not limited to, L-proline,trans-4-hydroxyl-L-proline, cis-4-hydroxyl-L-proline,trans-3-phenyl-L-proline, cis-3-phenyl-L-proline, L-(2-methyl)proline,L-pipecolinic acid, L-azetidine-2-carboxylic acid,L-indoline-2-carboxylic acid,L-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid,L-thiazolidine-4-carboxylic acid,L-(5,5-dimethyl)thiazolidine-4-carboxylic acid,L-thiamorpholine-3-carboxylic acid, glycine, 2-tert-butylglycine,D,L-phenylglycine, L-alanine, α-methylalanine, N-methyl-L-phenylalanine,L-diphenylalanine, sarcosine, D,L-phenylsarcosine, L-aspartic acidβ-tert-butyl ester, L-glutamic acid γ-tert-butyl ester,L-(O-benzyl)serine, 1-aminocyclopropanecarboxylic acid,1-aminocyclobutanecarboxylic acid, 1-aminocyclopentanecarboxylic acid(cycloleucine) 1-aminocyclohexanecarboxylic acid, L-serine and the like.If desired, the corresponding carboxylic acid esters of the amino acidsof Formula A, such as the methyl esters, ethyl esters and the like, canbe employed in the above reaction with the sulfonyl chloride B.Subsequent hydrolysis of the ester group to the carboxylic acid usingconventional reagents and conditions, i.e., treatment with an alkalimetal hydroxide in an inert diluent such as methanol/water, thenprovides the N-sulfonyl amino acid C.

Similarly, the sulfonyl chlorides of Formula B employed in the abovereaction are either known compounds or compounds that can be preparedfrom known compounds by conventional synthetic procedures. Suchcompounds are typically prepared from the corresponding sulfonic acid,i.e., from compounds of the formula R¹—SO₃H, using phosphoroustrichloride and phosphorous pentachloride. This reaction is generallyconducted by contacting the sulfonic acid with about 2 to 5 molarequivalents of phosphorous trichloride and phosphorous pentachloride,either neat or in an inert solvent, such as dichloromethane, attemperature in the range of about 0° C. to about 80° C. for about 1 toabout 48 hours to afford the sulfonyl chloride. Alternatively, thesulfonyl chlorides of Formula B can be prepared from the correspondingthiol compound, i.e., from compounds of the formula R¹—SH, by treatingthe thiol with chlorine (Cl₂) and water under conventional reactionconditions.

Examples of sulfonyl chlorides suitable for use in this inventioninclude, but are not limited to, methanesulfonyl chloride,2-propanesulfonyl chloride, 1-butanesulfonyl chloride, benzenesulfonylchloride, 1-naphthalenesulfonyl chloride, 2-naphthalenesulfonylchloride, p-toluenesulfonyl chloride, α-toluenesulfonyl chloride,4-acetamidobenzenesulfonyl chloride, 4-amidinobenzenesulfonyl chloride,4-tert-butylbenzenesulfonyl chloride, 4-bromobenzenesulfonyl chloride,2-carboxybenzenesulfonyl chloride, 4-cyanobenzenesulfonyl chloride,3,4-dichlorobenzenesulfonyl chloride, 3,5-dichlorobenzenesulfonylchloride, 3,4-dimethoxybenzenesulfonyl chloride,3,5-ditrifluoromethylbenzenesulfonyl chloride, 4-fluorobenzenesulfonylchloride, 4-methoxybenzenesulfonyl chloride,2-methoxycarbonylbenzenesulfonyl chloride, 4-methylamidobenzenesulfonylchloride, 4-nitrobenzenesulfonyl chloride, 4-thioamidobenzenesulfonylchloride, 4-trifluoromethylbenzenesulfonyl chloride,4-trifluoromethoxybenzenesulfonyl chloride,2,4,6-trimethylbenzenesulfonyl chloride, 2-phenylethanesulfonylchloride, 2-thiophenesulfonyl chloride, 5-chloro-2-thiophenesulfonylchloride, 2,5-dichloro-4-thiophenesulfonyl chloride, 2-thiazolesulfonylchloride, 2-methyl-4-thiazolesulfonyl chloride,1-methyl-4-imidazolesulfonyl chloride, 1-methyl-4-pyrazolesulfonylchloride, 5-chloro-1,3-dimethyl-4-pyrazolesulfonyl chloride,3-pyridinesulfonyl chloride, 2-pyrimidinesulfonyl chloride and the like.If desired, a sulfonyl fluoride, sulfonyl bromide or sulfonic acidanhydride may be used in place of the sulfonyl chloride in the abovereaction to form the N-sulfonyl amino acids of Formula C.

The intermediate N-sulfonyl amino acids of Formula C can also beprepared by reacting a sulfonamide of Formula D:

with a carboxylic acid derivative of the formula L(R³)CHCOOR^(y) where Lis a leaving group, such as chloro, bromo, iodo, mesylate, tosylate andthe like, and R^(y) is hydrogen or an alkyl group. This reaction istypically conducted by contacting the sulfonamide D with at least oneequivalent, preferably 1.1 to 2 equivalents, of the carboxylic acidderivative in the presence of a suitable base, such as triethylamine, inan inert diluent, such as DMF, at a temperature ranging from about 24°C. to about 37° C. for about 0.5 to about 4 hours. This reaction isfurther described in Zuckermann et al., J. Am. Chem. Soc., 1992, 114,10646-10647. Preferred carboxylic acid derivatives for use in thisreaction are α-chloro and α-bromocarboxylic acid esters such astert-butyl bromoacetate and the like. When a carboxylic acid ester isemployed in this reaction, the ester group is subsequently hydrolyzedusing conventional procedures to afford an N-sulfonyl amino acid ofFormula C.

The compounds of the present invention are then prepared by coupling theintermediate N-sulfonyl amino acid of Formula C with an amino acidderivative of Formula E:

This coupling reaction is typically conducted using well-known couplingreagents such as carbodiimides, BOP reagent(benzotriazol-1-yloxy-tris(dimethylamino)phosphoniumhexafluorophosphonate) and the like. Suitable carbodiimides include, byway of example, dicyclohexylcarbodiimide (DCC),1-(3-dimethylamino-propyl)-3-ethylcarbodiimide (EDC) and the like. Ifdesired, polymer supported forms of carbodiimide coupling reagents mayalso be used including, for example, those described in TetrahedronLetters, 34(48), 7685 (1993). Additionally, well-known couplingpromoters, such as N-hydroxysuccinimide, 1-hydroxybenzotriazole and thelike, may be used to facilitate the coupling reaction.

This coupling reaction is typically conducted by contacting theN-sulfonylamino acid C with about 1 to about 2 equivalents of thecoupling reagent and at least one equivalent, preferably about 1 toabout 1.2 equivalents, of amino acid derivative E in an inert diluent,such as dichloromethane, chloroform, acetonitrile, tetrahydrofuran,N,N-dimethylformamide and the like. Generally, this reaction isconducted at a temperature ranging from about 0° C. to about 37° C. forabout 12 to about 24 hours. Upon completion of the reaction, thecompound of the present invention is recovered by conventional methodsincluding neutralization, extraction, precipitation, chromatography,filtration, and the like.

Alternatively, the N-sulfonyl amino acid C can be converted into an acidhalide and the acid halide coupled with amino acid derivative E toprovide compounds of the present invention. The acid halide of C can beprepared by contacting C with an inorganic acid halide, such as thionylchloride, phosphorous trichloride, phosphorous tribromide or phosphorouspenta-chloride, or preferably, with oxalyl chloride under conventionalconditions. Generally, this reaction is conducted using about 1 to 5molar equivalents of the inorganic acid halide or oxalyl chloride,either neat or in an inert solvent, such as dichloromethane or carbontetrachloride, at temperature in the range of about 0° C. to about 80°C. for about 1 to about 48 hours. A catalyst, such as DMF, may also beused in this reaction.

The acid halide of N-sulfonyl amino acid C is then contacted with atleast one equivalent, preferably about 1.1 to about 1.5 equivalents, ofamino acid derivative E in an inert diluent, such as dichloromethane, ata temperature ranging from about −70° C. to about 40° C. for about 1 toabout 24 hours. Preferably, this reaction is conducted in the presenceof a suitable base to scavenge the acid generated during the reaction.Suitable bases include, by way of example, tertiary amines, such astriethylamine, diisopropylethylamine, N-methylmorpholine and the like.Alternatively, the reaction can be conducted under Schotten-Baumann-typeconditions using aqueous alkali, such as sodium hydroxide and the like.Upon completion of the reaction, the compound of the present inventionis recovered by conventional methods including neutralization,extraction, precipitation, chromatography, filtration, and the like.

Alternatively, the compounds of the present invention can be prepared byfirst forming a diamino acid derivative of Formula F:

The diamino acid derivatives of Formula F can be readily prepared bycoupling an amino acid of Formula A with an amino acid derivative ofFormula E using conventional amino acid coupling techniques andreagents, such carbodiimides, BOP reagent and the like, as describedabove. Diamino acid F can then be sulfonated using a sulfonyl chlorideof Formula B and using the synthetic procedures described above toprovide a compound of the present invention.

The amino acid derivatives of Formula E employed in the above reactionsare either known compounds or compounds that can be prepared from knowncompounds by conventional synthetic procedures. For example, amino acidderivatives of Formula E can be prepared by C-alkylating commerciallyavailable diethyl 2-acetamidomalonate (Aldrich, Milwaukee, Wis., USA)with an alkyl or substituted alkyl halide. This reaction is typicallyconducted by treating the diethyl 2-acetamidomalonate with at least oneequivalent of sodium ethoxide and at least one equivalent of an alkyl orsubstituted alkyl halide in refluxing ethanol for about 6 to about 12hours. The resulting C-alkylated malonate is then deacetylated,hydrolyzed and decarboxylated by heating in aqueous hydrochloric acid atreflux for about 6 to about 12 hours to provide the amino acid,typically as the hydrochloride salt.

Examples of amino acid derivatives of Formula E suitable for use in theabove reactions include, but are not limited to, L-tyrosine methylester, L-3,5-diiodotyrosine methyl ester, L-3-iodotyrosine methyl ester,β-(4-hydroxy-naphth-1-yl)-L-alanine methyl ester,β-(6-hydroxy-naphth-2-yl)-L-alanine methyl ester, and the like. Ifdesired, of course, other esters or amides of the above-describedcompounds may also be employed.

For ease of synthesis, the compounds of the present invention aretypically prepared as an ester, i.e., where R⁶ is an alkoxy orsubstituted alkoxy group and the like. If desired, the ester group canbe hydrolysed using conventional conditions and reagents to provide thecorresponding carboxylic acid. Typically, this reaction is conducted bytreating the ester with at least one equivalent of an alkali metalhydroxide, such as lithium, sodium or potassium hydroxide, in an inertdiluent, such as methanol or mixtures of methanol and water, at atemperature ranging about 0° C. to about 24° C. for about 1 to about 10hours. Alternatively, benzyl esters may be removed by hydrogenolysisusing a palladium catalyst, such as palladium on carbon. The resultingcarboxylic acids may be coupled, if desired, to amines such as β-alanineethyl ester, hydroxyamines such as hydroxylamine andN-hydroxysuccinimide, alkoxyamines and substituted alkoxyamines such asO-methylhydroxylamine and O-benzylhydroxylamine, and the like, usingconventional coupling reagents and conditions as described above.

As will be apparent to those skilled in the art, other functional groupspresent on any of the substituents of the compounds of the presentinvention can be readily modified or derivatized either before or afterthe above-described coupling reactions using well-known syntheticprocedures. For example, a nitro group present on a substituent of acompound of the present invention or an intermediate thereof may bereadily reduced by hydrogenation in the presence of a palladiumcatalyst, such as palladium on carbon, to provide the correspondingamino group. This reaction is typically conducted at a temperature offrom about 20° C. to about 50° C. for about 6 to about 24 hours in aninert diluent, such as methanol. Compounds having a nitro group on,e.g., the R³ substituent, can be prepared, for example, by using a4-nitrophenylalanine derivative and the like in the above-describedcoupling reactions.

Similarly, a pyridyl group can be hydrogenated in the presence of aplatinum catalyst, such as platinum oxide, in an acidic diluent toprovide the corresponding piperidinyl analogue. Generally, this reactionis conducted by treating the pyridine compound with hydrogen at apressure ranging from about 20 psi to about 60 psi, preferably about 40psi, in the presence of the catalyst at a temperature of about 20° C. toabout 50° C. for about 2 to about 24 hours in an acidic diluent, such asa mixture of methanol and aqueous hydrochloric acid. Compounds having apyridyl group can be readily prepared by using, for example,β-(2-pyridyl)-, β-(3-pyridyl)- or β-(4-pyridyl)-L-alanine derivatives inthe above-described coupling reactions.

Additionally, when a substituent of a compound of the present inventionor an intermediate thereof contains a primary or secondary amino group,such amino groups can be further derivatized either before or after theabove coupling reactions to provide, by way of example, amides,sulfonamides, ureas, thioureas, carbamates, secondary or tertiary aminesand the like. Compounds having a primary amino group on such asubstituent may be prepared, for example, by reduction of thecorresponding nitro compound as described above. Alternatively, suchcompounds can be prepared by using an amino acid derivative of Formula Ederived from lysine, 4-aminophenylalanine and the like in theabove-described coupling reactions.

By way of illustration, a compound of the present invention or anintermediate thereof having a substituent containing a primary orsecondary amino group can be readily N-acylated using conventionalacylating reagents and conditions to provide the corresponding amide.This acylation reaction is typically conducted by treating the aminocompound with at least one equivalent, preferably about 1.1 to about 1.2equivalents, of a carboxylic acid in the presence of a coupling reagentsuch as a carbodiimide, BOP reagent(benzotriazol-1-yloxy-tris(dimethylamino)-phosphoniumhexafluorophosphonate) and the like, in an inert diluent, such asdichloromethane, chloroform, acetonitrile, tetrahydrofuran,N,N-dimethylformamide and the like, at a temperature ranging from about0° C. to about 37° C. for about 4 to about 24 hours. Preferably, apromoter, such as N-hydroxysuccinimide, 1-hydroxybenzotriazole and thelike, is used to facilitate the acylation reaction. Examples ofcarboxylic acids suitable for use in this reaction include, but are notlimited to, N-tert-butyloxycarbonylglycine,N-tert-butyloxycarbonyl-L-phenylalanine,N-tert-butyloxycarbonyl-L-aspartic acid benzyl ester, benzoic acid,N-tert-butyloxycarbonylisonipecotic acid, N-methylisonipecotic acid,N-tert-butyloxycarbonylnipecotic acid,N-tert-butyloxycarbonyl-L-tetrahydroisoquinoline-3-carboxylic acid,N-(toluene-4-sulfonyl)-L-proline and the like.

Alternatively, a compound of the present invention or an intermediatethereof containing a primary or secondary amino group can be N-acylatedusing an acyl halide or a carboxylic acid anhydride to form thecorresponding amide. This reaction is typically conducted by contactingthe amino compound with at least one equivalent, preferably about 1.1 toabout 1.2 equivalents, of the acyl halide or carboxylic acid anhydridein an inert diluent, such as dichloromethane, at a temperature rangingfrom about of about −70° C. to about 40° C. for about 1 to about 24hours. If desired, an acylation catalyst such as4-(N,N-dimethylamino)pyridine may be used to promote the acylationreaction. The acylation reaction is preferably conducted in the presenceof a suitable base to scavenge the acid generated during the reaction.Suitable bases include, by way of example, tertiary amines, such astriethylamine, diisopropylethylamine, N-methylmorpholine and the like.Alternatively, the reaction can be conducted under Schotten-Baumann-typeconditions using aqueous alkali, such as sodium hydroxide and the like.

Examples of acyl halides and carboxylic acid anhydrides suitable for usein this reaction include, but are not limited to, 2-methylpropionylchloride, trimethylacetyl chloride, phenylacetyl chloride, benzoylchloride, 2-bromobenzoyl chloride, 2-methylbenzoyl chloride,2-trifluoromethylbenzoyl chloride, isonicotinoyl chloride, nicotinoylchloride, picolinoyl chloride, acetic anhydride, succinic anhydride andthe like. Carbamyl chlorides, such as N,N-dimethylcarbamyl chloride,N,N-diethylcarbamyl chloride and the like, can also be used in thisreaction to provide ureas. Similarly, dicarbonates, such asdi-tert-butyl dicarbonate, may be employed to provide carbamates.

In a similar manner, a compound of the present invention or anintermediate thereof containing a primary or secondary amino group maybe N-sulfonated to form a sulfonamide using a sulfonyl halide orsulfonic acid anhydride. Sulfonyl halides and sulfonic acid anhydridessuitable for use in this reaction include, but are not limited to,methanesulfonyl chloride, chloromethanesulfonyl chloride,p-toluenesulfonyl chloride, trifluoromethanesulfonic anhydride and thelike. Similarly, sulfamoyl chlorides, such as dimethylsulfamoylchloride, can be used to provide sulfamides (e.g., >N—SO₂—N<).

Additionally, a primary and secondary amino group present on asubstituent of a compound of the present invention or an intermediatethereof can be reacted with an isocyanate or a thioisocyanate to give aurea or thiourea, respectively. This reaction is typically conducted bycontacting the amino compound with at least one equivalent, preferablyabout 1.1 to about 1.2 equivalents, of the isocyanate or thioisocyanatein an inert diluent, such as toluene and the like, at a temperatureranging from about 24° C. to about 37° C. for about 12 to about 24hours. The isocyanates and thioisocyanates used in this reaction arecommercially available or can be prepared from commercially availablecompounds using well-known synthetic procedures. For example,isocyanates and thioisocyanates are readily prepared by reacting theappropriate amine with phosgene or thiophosgene. Examples of isocyanatesand thioisocyanates suitable for use in this reaction include, but arenot limited to, ethyl isocyanate, n-propyl isocyanate, 4-cyanophenylisocyanate, 3-methoxyphenyl isocyanate, 2-phenylethyl isocyanate, methylthioisocyanate, ethyl thioisocyanate, 2-phenylethyl thioisocyanate,3-phenylpropyl thioisocyanate, 3-(N,N-diethylamino)propylthioisocyanate, phenyl thioisocyanate, benzyl thioisocyanate, 3-pyridylthioisocyanate, fluorescein isothiocyanate (isomer L), and the like.

Furthermore, when a compound of the present invention or an intermediatethereof contains a primary or secondary amino group, the amino group canbe reductively alkylated using aldehydes or ketones to form a secondaryor tertiary amino group. This reaction is typically conducted bycontacting the amino compound with at least one equivalent, preferablyabout 1.1 to about 1.5 equivalents, of an aldehyde or ketone and atleast one equivalent based on the amino compound of a metal hydridereducing agent, such as sodium cyanoborohydride, in an inert diluent,such as methanol, tetrahydrofuran, mixtures thereof and the like, at atemperature ranging from about 0° C. to about 50° C. for about 1 toabout 72 hours. Aldehydes and ketones suitable for use in this reactioninclude, by way of example, benzaldehyde, 4-chlorobenzaldehyde,valeraldehyde and the like.

In a similar manner, when a compound of the present invention or anintermediate thereof has a substituent containing a hydroxyl group, thehydroxyl group can be further modified or derivatized either before orafter the above coupling reactions to provide, by way of example,ethers, carbamates and the like. Compounds of Formulae I and II having ahydroxyl group on the R⁵ substituent, for example, can be prepared usingan amino acid derivative of Formula E derived from tyrosine and the likein the above-described reactions.

By way of example, a compound of the present invention or anintermediate thereof having a substituent containing a hydroxyl groupcan be readily O-alkylated to form ethers. This O-alkylation reaction istypically conducted by contacting the hydroxy compound with a suitablealkali or alkaline earth metal base, such as potassium carbonate, in aninert diluent, such as acetone, 2-butanone and the like, to form thealkali or alkaline earth metal salt of the hydroxyl group. This salt isgenerally not isolated, but is reacted in situ with at least oneequivalent of an alkyl or substituted alkyl halide or sulfonate, such asan alkyl chloride, bromide, iodide, mesylate or tosylate, to afford theether. Generally, this reaction is conducted at a temperature rangingfrom about 60° C. to about 150° C. for about 24 to about 72 hours.Preferably, a catalytic amount of sodium or potassium iodide is added tothe reaction mixture when an alkyl chloride or bromide is employed inthe reaction.

Examples of alkyl or substituted alkyl halides and sulfonates suitablefor use in this reaction include, but are not limited to, tert-butylbromoacetate, N-tert-butyl chloroacetamide, 1-bromoethylbenzene, ethylα-bromophenylacetate, 2-(N-ethyl-N-phenylamino)ethyl chloride,2-(N,N-ethylamino)ethyl chloride, 2-(N,N-diisopropylamino)ethylchloride, 2-(N,N-dibenzylamino)ethyl chloride, 3-(N,N-ethylamino)propylchloride, 3-(N-benzyl-N-methylamino)propyl chloride,N-(2-chloroethyl)morpholine, 2-(hexamethyleneimino)ethyl chloride,3-(N-methylpiperazine)propyl chloride,1-(3-chlorophenyl)-4-(3-chloropropyl)piperazine,2-(4-hydroxy-4-phenylpiperidine)ethyl chloride,N-tert-butyloxycarbonyl-3-piperidinemethyl tosylate and the like.

Alternatively, a hydroxyl group present on a substituent of a compoundof the present invention or an intermediate thereof can be O-alkylatingusing the Mitsunobu reaction. In this reaction, an alcohol, such as3-(N,N-dimethylamino)-1-propanol and the like, is reacted with about 1.0to about 1.3 equivalents of triphenylphosphine and about 1.0 to about1.3 equivalents of diethyl azodicarboxylate in an inert diluent, such astetrahydrofuran, at a temperature ranging from about −10° C. to about 5°C. for about 0.25 to about 1 hour. About 1.0 to about 1.3 equivalents ofa hydroxy compound, such as N-tert-butyltyrosine methyl ester, is thenadded and the reaction mixture is stirred at a temperature of about 0°C. to about 30° C. for about 2 to about 48 hours to provide theO-alkylated product.

In a similar manner, a compound of the present invention or anintermediate thereof containing a aryl hydroxy group can be reacted withan aryl iodide to provide a diaryl ether. Generally, this reaction isconducted by forming the alkali metal salt of the hydroxyl group using asuitable base, such as sodium hydride, in an inert diluent such asxylenes at a temperature of about −25° C. to about 10° C. The salt isthen treated with about 1.1 to about 1.5 equivalents of cuprous bromidedimethyl sulfide complex at a temperature ranging from about 10° C. toabout 30° C. for about 0.5 to about 2.0 hours, followed by about 1.1 toabout 1.5 equivalents of an aryl iodide, such as sodium 2-iodobenzoateand the like. The reaction is then heated to about 70° C. to about 150°C. for about 2 to about 24 hours to provide the diaryl ether.

Additionally, a hydroxy-containing compound can also be readilyderivatized to form a carbamate. In one method for preparing suchcarbamates, a hydroxy compound of the present invention or anintermediate thereof is contacted with about 1.0 to about 1.2equivalents of 4-nitrophenyl chloroformate in an inert diluent, such asdichloromethane, at a temperature ranging from about −25° C. to about 0°C. for about 0.5 to about 2.0 hours. Treatment of the resultingcarbonate with an excess, preferably about 2 to about 5 equivalents, ofa trialkylamine, such as triethylamine, for about 0.5 to 2 hours,followed by about 1.0 to about 1.5 equivalents of a primary or secondaryamine provides the carbamate. Examples of amines suitable for using inthis reaction include, but are not limited to, piperazine,1-methylpiperazine, 1-acetylpiperazine, morpholine, thiomorpholine,pyrrolidine, piperidine and the like.

Alternatively, in another method for preparing carbamates, ahydroxy-containing compound is contacted with about 1.0 to about 1.5equivalents of a carbamyl chloride in an inert diluent, such asdichloromethane, at a temperature ranging from about 25° C. to about 70°C. for about 2 to about 72 hours. Typically, this reaction is conductedin the presence of a suitable base to scavenge the acid generated duringthe reaction. Suitable bases include, by way of example, tertiaryamines, such as triethylamine, diisopropylethylamine, N-methylmorpholineand the like. Additionally, at least one equivalent (based on thehydroxy compound) of 4-(N,N-dimethylamino)pyridine is preferably addedto the reaction mixture to facilitate the reaction. Examples of carbamylchlorides suitable for use in this reaction include, by way of example,dimethylcarbamyl chloride, diethylcarbamyl chloride and the like.

Likewise, when a compound of the present invention or an intermediatethereof contains a primary or secondary hydroxyl group, such hydroxylgroups can be readily converted into a leaving group and displaced toform, for example, amines, sulfides and fluorides. For example,derivatives of 4-hydroxy-L-proline can be converted into thecorresponding 4-amino, 4-thio or 4-fluoro-L-proline derivatives vianucleophilic displacement of the derivatized hydroxyl group. Generally,when a chiral compound is employed in these reactions, thestereochemistry at the carbon atom attached to the derivatized hydroxylgroup is typically inverted.

These reactions are typically conducted by first converting the hydroxylgroup into a leaving group, such as a tosylate, by treatment of thehydroxy compound with at least one equivalent of a sulfonyl halide, suchas p-toluenesulfonyl chloride and the like, in pyridine. This reactionis generally conducted at a temperature of from about 0° C. to about 70°C. for about 1 to about 48 hours. The resulting tosylate can then bereadily displaced with sodium azide, for example, by contacting thetosylate with at least one equivalent of sodium azide in an inertdiluent, such as a mixture of N,N-dimethylformamide and water, at atemperature ranging from about 0° C. to about 37° C. for about 1 toabout 12 hours to provide the corresponding azido compound. The azidogroup can then be reduced by, for example, hydrogenation using apalladium on carbon catalyst to provide the amino (—NH₂) compound.

Similarly, a tosylate group can be readily displaced by a thiol to forma sulfide. This reaction is typically conducted by contacting thetosylate with at least one equivalent of a thiol, such as thiophenol, inthe presence of a suitable base, such as1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), in an inert diluent, such asN,N-dimethylformamide, at a temperature of from about 0° C. to about 37°C. for about 1 to about 12 hours to provide the sulfide. Additionally,treatment of a tosylate with morpholinosulfur trifluoride in an inertdiluent, such as dichloromethane, at a temperature ranging from about 0°C. to about 37° C. for about 12 to about 24 hours affords thecorresponding fluoro compound.

Furthermore, a compound of the present invention or an intermediatethereof having a substituent containing an iodoaryl group, for example,when R⁵, of Formula I or II, is a (4-iodophenyl)methyl group, can bereadily converted either before or after the above coupling reactionsinto a biaryl compound. Typically, this reaction is conducted bytreating the iodoaryl compound with about 1.1 to about 2 equivalents ofan arylzinc iodide, such as 2-(methoxycarbonyl)phenylzinc iodide, in thepresence of a palladium catalyst, such as palladiumtetra(triphenylphosphine), in an inert diluent, such as tetrahydrofuran,at a temperature ranging from about 24° C. to about 30° C. until thereaction is complete. This reaction is further described, for example,in Rieke, J. Org. Chem. 1991, 56, 1445.

In some cases, the compounds of the present invention or intermediatesthereof may contain substituents having one or more sulfur atoms. Suchsulfur atoms will be present, for example, when the amino acid ofFormula A employed in the above reactions is derived fromL-thiazolidine-4-carboxylic acid,L-(5,5-dimethyl)thiazolidine-4-carboxylic acid,L-thiamorpholine-3-carboxylic acid and the like. When present, suchsulfur atoms can be oxidized either before or after the above couplingreactions to provide a sulfoxide or sulfone compound using conventionalreagents and reaction conditions. Suitable reagents for oxidizing asulfide compound to a sulfoxide include, by way of example, hydrogenperoxide, 3-chloroperoxybenzoic acid (MCPBA), sodium periodate and thelike. The oxidation reaction is typically conducted by contacting thesulfide compound with about 0.95 to about 1.1 equivalents of theoxidizing reagent in an inert diluent, such as dichloromethane, at atemperature ranging from about −50° C. to about 75° C. for about 1 toabout 24 hours. The resulting sulfoxide can then be further oxidized tothe corresponding sulfone by contacting the sulfoxide with at least oneadditional equivalent of an oxidizing reagent, such as hydrogenperoxide, MCPBA, potassium permanganate and the like. Alternatively, thesulfone can be prepared directly by contacting the sulfide with at leasttwo equivalents, and preferably an excess, of the oxidizing reagent.Such reactions are described further in March, “Advanced OrganicChemistry”, 4th Ed., pp. 1202-1202, Wiley Publishers, (1992).

As described above, the compounds of the present invention having an R²substituent other an hydrogen can be prepared using an N-substitutedamino acid of Formula A, such as sarcosine, N-methyl-L-phenylalanine andthe like, in the above-described coupling reactions. Alternatively, suchcompounds can be prepared by N-alkylation of a sulfonamide of Formula Ior C (where R² is hydrogen) using conventional synthetic procedures.Typically, this N-alkylation reaction is conducted by contacting thesulfonamide with at least one equivalent, preferably 1.1 to 2equivalents, of an alkyl or substituted alkyl halide in the presence ofa suitable base, such as potassium carbonate, in an inert diluent, suchas acetone, 2-butanone and the like, at a temperature ranging from about25° C. to about 70° C. for about 2-to about 48 hours. Examples of alkylor substituted alkyl halides suitable for use in this reaction include,but are not limited to, methyl iodide, and the like.

Additionally, the sulfonamides of Formula I or C wherein R² is hydrogenand R′is a 2-alkoxycarbonylaryl group can be intramolecularly cyclizedto form 1,2-benzisothiazol-3-one derivatives or analogues thereof. Thisreaction is typically conducted by treating a sulfonamide, such asN-(2-methoxycarbonylphenylsulfonyl)glycine-L-phenylalanine benzyl ester,with about 1.0 to 1.5 equivalents of a suitable base, such as an alkalimetal hydride, in a inert diluent, such as tetrahydrofuran, at atemperature ranging from about 0° C. to about 30° C. for about 2 toabout 48 hours to afford the cyclized 1,2-benzisothiazol-3-onederivative.

Lastly, the compounds of Formula I or II where Q is —C(S)NR⁷— areprepared by using an amino thionoacid derivative in place of amino acidA in the above described synthetic procedures. Such amino thionoacidderivatives can be prepared by the procedures described in Shalaky etal., J. Org. Chem., 61:9045-9048 (1996) and Brain et al., J. Org. Chem.,62:3808-3809 (1997) and references cited therein.

Compounds of Formulae III-IX

In one aspect, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compounds ofFormulae IIIa, IIIb, IVa, IVb, Va, Vb, Vc, Vd, Ve, VIa, VIb, VIIa, VIIb,VIIc, VIId, VIIe, VIIIa, VIIIb, IXa, IXb, IXc, IXd, and IXe.

In one aspect, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compoundsdefined by Formula IIIa and/or IIIb below. These compounds have abinding affinity to VLA-4 as expressed by an IC₅₀ of about 15 μM or less(measured as described in Example A below):

-   -   wherein, in Formula IIIa, R¹ and R², together with the carbon        atom and W to which they are bound respectively, are joined to        form an aryl, cycloalkenyl, heteroaryl or heterocyclic group        having at least five atoms in the aryl, cycloalkenyl, heteroaryl        or heterocyclic group and optionally containing or additionally        containing in the case of heteroaryl and heterocyclic groups 1        to 3 heteroatoms selected from the group consisting of oxygen,        nitrogen and sulfur, and wherein the heteroaryl or heterocyclic        group is mono-cyclic;    -   in Formula IIIb, R¹ and R², together with the carbon atom and W′        to which they are bound respectively, are joined to form a        cycloalkyl, cycloalkenyl or heterocyclic group having at least        five atoms in the cycloalkyl, cycloalkenyl or heterocyclic group        and optionally containing or additionally containing in the case        of the heterocyclic group 1 to 3 heteroatoms selected from the        group consisting of oxygen, nitrogen and sulfur, and wherein the        heterocyclic group is mono-cyclic;    -   and further wherein said aryl, cycloalkyl, cycloalkenyl,        heteroaryl or heterocyclic group of Formula IIIa or IIIb is        optionally substituted, on any ring atom capable of        substitution, with 1-3 substituents selected from the group        consisting of alkyl, substituted alkyl, alkoxy, substituted        alkoxy, acyl, acylamino, thiocarbonylamino, acyloxy, amino,        substituted amino, amidino, alkyl amidino, thioamidino,        aminoacyl, aminocarbonylamino, aminothiocarbonylamino,        aminocarbonyloxy, aryl, substituted aryl, aryloxy, substituted        aryloxy, aryloxyaryl, substituted aryloxyaryl, cyano, halogen,        hydroxyl, nitro, oxo, carboxyl, cycloalkyl, substituted        cycloalkyl, guanidino, guanidinosulfone, thiol, thioalkyl,        substituted thioalkyl, thioaryl, substituted thioaryl,        thiocycloalkyl, substituted thiocycloalkyl, thioheteroaryl,        substituted thioheteroaryl, thioheterocyclic, substituted        thioheterocyclic, heteroaryl, substituted heteroaryl,        heterocyclic, substituted heterocyclic, cycloalkoxy, substituted        cycloalkoxy, heteroaryloxy, substituted heteroaryloxy,        heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino,        oxythiocarbonylamino, —OS(O)₂-alkyl, —OS(O)₂-substituted alkyl,        —OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,        —OS(O)₂-substituted heteroaryl, —OS(O)₂-heterocyclic,        —OS(0)₂-substituted heterocyclic, —OSO₂—NRR where each R is        independently hydrogen or alkyl, —NRS (O)₂-alkyl,        —NRS(O)₂-substituted alkyl, —NRS(O)₂-aryl, —NRS(O)₂-substituted        aryl, —NRS(O)₂-heteroaryl, —NRS(O)₂-substituted heteroaryl,        —NRS(O)₂-heterocyclic, —NRS(O)₂-substituted heterocyclic,        —NRS(O)₂—NR-alkyl, —NRS(O)₂—NR-substituted alkyl,        —NRS(O)₂—NR-aryl, —NRS(O)₂—NR-substituted aryl,        —NRS(O)₂—NR-heteroaryl, —NRS(O)₂—NR-substituted heteroaryl,        —NRS(O)₂—NR-heterocyclic, —NRS(O)₂—NR-substituted heterocyclic        where R is hydrogen or alkyl, —N[S(O)₂—R′]₂ and —N[S(O)₂—NR′9 ₂        where each R′ is independently selected from the group        consisting of alkyl, substituted alkyl, aryl, substituted aryl,        heteroaryl, substituted heteroaryl, heterocyclic and substituted        heterocyclic;    -   R³ and R^(3′) are independently selected from the group        consisting of hydrogen, isopropyl, —CH₂Z where Z is selected        from the group consisting of hydrogen, hydroxyl, acylamino,        alkyl, alkoxy, aryloxy, aryl, aryloxyaryl, carboxyl,        carboxylalkyl, carboxyl-substituted alkyl, carboxyl-cycloalkyl,        carboxyl-substituted cycloalkyl, carboxylaryl,        carboxyl-substituted aryl, carboxylheteroaryl,        carboxyl-substituted heteroaryl, carboxylheterocyclic,        carboxyl-substituted heterocyclic, cycloalkyl, substituted        alkyl, substituted alkoxy, substituted aryl, substituted        aryloxy, substituted aryloxyaryl, substituted cycloalkyl,        heteroaryl, substituted heteroaryl, heterocyclic and substituted        heterocyclic, and    -   where R³ and R^(3′) are joined to form a substituent selected        from the group consisting of ═CHZ where Z is defined above        provided that Z is not hydroxyl or thiol, cycloalkyl,        substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl,        heterocyclic and substituted heterocyclic;    -   Q is selected from the group consisting of —O—, —S—, —S(O)—,        —S(O)₂, and —NR⁴—;    -   R⁴ is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, alkenyl, substituted alkenyl, cycloalkyl,        substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl,        aryl, substituted aryl, heteroaryl, substituted heteroaryl,        heterocyclic and substituted heterocyclic or, optionally, R⁴ and        R¹ or R⁴ and R², together with the atoms to which they are        bound, are joined to form a heteroaryl, a substituted        heteroaryl, a heterocyclic or a substituted heterocyclic group;    -   W is selected from the group consisting of nitrogen and carbon;        and    -   W′ is selected from the group consisting of nitrogen, carbon,        oxygen, sulfur, S(O), and S(O)₂;    -   X is selected from the group consisting of hydroxyl, alkoxy,        substituted alkoxy, alkenoxy, substituted alkenoxy, cycloalkoxy,        substituted cycloalkoxy, cycloalkenoxy, substituted        cycloalkenoxy, aryloxy, substituted aryloxy, heteroaryloxy,        substituted heteroaryloxy, heterocyclyloxy, substituted        heterocyclyloxy and —NR″R′ where each R″ is independently        selected from the group consisting of hydrogen, alkyl,        substituted alkyl, alkenyl, substituted alkenyl, cycloalkyl,        substituted cycloalkyl, aryl, substituted aryl, heteroaryl,        substituted heteroaryl, heterocyclic and substituted        heterocyclic;    -   and enantiomers, diastereomers and pharmaceutically acceptable        salts thereof;    -   and further wherein the compound of Formula IIIa and/or IIIb has        a binding affinity to VLA-4 as expressed by an IC₅₀ of about 15        μM or less.

Preferably, R³ is —(CH₂)_(x)—Ar—R⁹, where Ar is aryl, substituted aryl,heteroaryl and substituted heteroaryl; R⁹ is selected from the groupconsisting of acyl, acylamino, acyloxy, aminoacyl, aminocarbonylamino,aminothiocarbonylamino, aminocarbonyloxy, oxythiocarbonylamino,thioamidino, thiocarbonylamino, aminosulfonylamino, aminosulfonyloxy,aminosulfonyl, oxysulfonylamino and oxysulfonyl; and x is an integerfrom 0 to 4. R^(3′) is preferably alkyl or hydrogen; more preferably, R³is hydrogen.

More preferably, R³is a group of the formula:

-   -   wherein R⁹ and x are as defined herein. Preferably, R⁹ is in        thepara position of the phenyl ring; and x is an integer of from        1 to 4, more preferably, x is 1.

In a preferred embodiment, R⁹ is selected from —O—Z—NR¹¹R^(11′) and—O—Z—R¹² wherein R¹¹ and R^(11′) are independently selected from thegroup consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl,heterocyclic, substituted heterocyclic, and where R¹¹ and R^(11′) arejoined to form a heterocycle or a substituted heterocycle, R¹² isselected from the group consisting of heterocycle and substitutedheterocycle, and Z is selected from the group consisting of —C(O)— and—SO₂—. More preferably, R⁹ is —OC(O)NR¹¹R^(11′), wherein R¹¹ and R^(11′)are as defined herein.

Z is preferably —C(O)—. Preferably, Q is —NR⁴—.

In a preferred embodiment, the compounds that can be utilized ascombination therapies with methotrexate for the treatment of RA arecompounds defined by Formula IVa and/or IVb below.

-   -   wherein R³, R^(3′) and X are as defined herein;    -   ring A and ring B independently form a heteroaryl or substituted        heteroaryl group having two nitrogen atoms in the heteroaryl        ring;    -   R⁵ is selected from the group consisting of alkyl, substituted        alkyl, alkenyl, substituted alkenyl, aryl, substituted aryl,        cycloalkyl, substituted cycloalkyl, cycloalkeny, substituted        cycloalkenyl, heterocyclic, substituted heterocylic, heteroaryl        and substituted heteroaryl;    -   R⁶ is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, cycloalkyl, substituted cycloalkyl,        cycloalkenyl, substituted cycloalkenyl, heterocyclic,        substituted heterocyclic, aryl, substituted aryl, heteroaryl,        substituted heteroaryl, and —SO₂R¹⁰ where R¹⁰ is selected from        the group consisting of alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl,        heterocyclic, substituted heterocyclic, aryl, substituted aryl,        heteroaryl, substituted heteroaryl;    -   or optionally, one of, R⁴ and ring A, R⁴ and R⁵, R⁴ and R⁵ and        R⁶, together with the atoms to which they are bound, can be        joined to form a hetereocyclic or substituted heterocyclic ring;    -   and enantiomers, diastereomers and pharmaceutically acceptable        salts thereof; and provided that ring B does not form a 6-amino        or substituted amino pyrimidin-4-yl group.

Preferably, ring A forms a pyridazine, pyrimidine or pyrazine ring; morepreferably, a pyrimidine or pyrazine ring; wherein the pyridazine,pyrimidine or pyrazine ring is optionally substituted with 1 to 3substituents selected from the group consisting of alkyl, substitutedalkyl, alkoxy, substituted alkoxy, amino, substituted amino, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic and halogen.

Preferably, ring B forms a pyridazine, pyrimidine, pyrazine,1-oxo-1,2,5-thiadiazole or a 1,1-dioxo-1,2,5-thiadiazole ring; morepreferably, a pyrimidine, pyrazine, 1-oxo-1,2,5-thiadiazole or a1,1-dioxo-1,2,5-thiadiazole ring; wherein the pyridazine, pyrimidine orpyrazine ring is optionally substituted with 1 to 3 substituentsselected from the group consisting of alkyl, substituted alkyl, alkoxy,substituted alkoxy, amino, substituted amino, cycloalkyl, substitutedcycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic, substituted heterocyclic and halogen.

Preferably the compounds that can be utilized as combination therapieswith methotrexate for the treatment of RA are compounds defined byFormula IVa.

In another preferred embodiment, the compounds that can be utilized ascombination therapies with methotrexate for the treatment of RA arecompounds defined by Formula Va, Vb, Vc, Vd, or Ve:

-   -   wherein R³, R^(3′) and X are as defined herein;    -   R^(4′) is selected from the group consisting of hydrogen and        alkyl or, optionally, one of, R^(4′) and R⁵, R^(4′) and R⁶, R⁵        and R⁶, R⁵ and R⁸, or R⁶ and R⁸, together with the atoms to        which they are bound, are joined to form a heterocyclic, a        substituted heterocyclic, a heteroaryl or substituted heteroaryl        group optionally containing from 1 to 3 additional hetero ring        atoms selected from the group consisting of oxygen, nitrogen and        sulfur;    -   R^(4′) is selected from the group consisting of hydrogen and        alkyl;    -   R⁵ is selected from the group consisting of alkyl, substituted        alkyl, alkenyl, substituted alkenyl, aryl, substituted aryl,        cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted        cycloalkenyl, heterocyclic, substituted heterocylic, heteroaryl        and substituted heteroaryl;    -   R⁶ is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, cycloalkyl, substituted cycloalkyl,        cycloalkenyl, substituted cycloalkenyl, heterocyclic,        substituted heterocyclic, aryl, substituted aryl, heteroaryl,        substituted heteroaryl, and —SO₂R¹⁰ where R¹⁰ is selected from        the group consisting of alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl,        heterocyclic, substituted heterocyclic, aryl, substituted aryl,        heteroaryl, substituted heteroaryl;    -   R⁷ and R⁸ are independently selected from the group consisting        of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted        cycloalkyl, amino, substituted amino, aryl, substituted aryl,        heteroaryl, substituted heteroaryl, heterocyclic, substituted        heterocyclic and halogen;    -   R¹⁶ and R¹⁷ are independently selected from the group consisting        of hydrogen, alkyl, substituted alkyl, alkoxy, substituted        alkoxy, amino, substituted amino, cycloalkyl, substituted        cycloalkyl, aryl, substituted aryl, heteroaryl, substituted        heteroaryl, heterocyclic, substituted heterocyclic and halogen;        and    -   R¹⁸ is selected from the group consisting of alkyl, substituted        alkyl, alkoxy, substituted alkoxy, amino, substituted amino,        cycloalkyl, substituted cycloalkyl, aryl, substituted aryl,        heteroaryl, substituted heteroaryl, heterocyclic and substituted        heterocyclic;    -   R²⁰ is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, alkoxy, substituted alkoxy, cycloalkyl,        substituted cycloalkyl, aryl, substituted aryl, heteroaryl,        substituted heteroaryl, heterocyclic, substituted heterocyclic        and halogen;    -   R²¹ is selected from the group consisting of alkyl, substituted        alkyl, alkoxy, substituted alkoxy, amino, substituted amino,        cycloalkyl, substituted cycloalkyl, aryl, substituted aryl,        heterocyclic and substituted heterocyclic;    -   b is 1 or 2;    -   and enantiomers, diastereomers and pharmaceutically acceptable        salts thereof.

Preferably, the compounds that can be utilized as combination therapieswith methotrexate for the treatment of RA are compounds of Formula Va,Vc, or Vd.

Preferably, in the compounds of Formula Va, Vc, and Vd

-   -   R⁵, R¹⁸, and R²¹ are independently selected from the group        consisting of heteroaryl, substituted heteroaryl, heterocyclic,        and substituted heterocyclic;    -   R⁷ and R¹⁶ are independently amino or substituted amino;    -   R³ is a group of the formula:    -   wherein R⁹ is selected from the group consisting acyl,        acylamino, acyloxy, aminoacyl, aminocarbonylamino,        aminothiocarbonylamino, aminocarbonyloxy, oxycarbonylamino,        oxythiocarbonylamino, thioamidino, thiocarbonylamino,        aminosulfonylamino, aminosulfonyloxy, aminosulfonyl,        oxysulfonylamino and oxysulfonyl; and x is an integer from 0 to        4; and    -   R³ is hydrogen.

In another embodiment, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compoundsdefined by Formula VIa and/or VIb:

-   -   wherein, in Formula VIa, R¹ and R², together with the carbon        atom and W to which they are bound respectively, are joined to        form an aryl, cycloalkenyl, heteroaryl or heterocyclic group        having at least five atoms in the aryl, cycloalkenyl, heteroaryl        or heterocyclic group and optionally containing or additionally        containing in the case of heteroaryl and heterocyclic groups 1        to 3 heteroatoms selected from the group consisting of oxygen,        nitrogen and sulfur, and wherein the heteroaryl or heterocyclic        group is mono-cyclic;    -   in Formula VIb, R¹ and R², together with the carbon atom and W′        to which they are bound respectively, are joined to form a        cycioalkyl, cycloalkenyl or heterocyclic group having at least        five atoms in the cycloalkyl, cycloalkenyl or heterocyclic group        and optionally containing or additionally containing in the case        the heterocyclic group 1 to 3 heteroatoms selected from the        group consisting of oxygen, nitrogen and sulfur, and wherein the        heterocyclic group is mono-cyclic;    -   and further wherein said aryl, cycloalkyl, cycloalkenyl,        heteroaryl or heterocyclic group of Formula VIa or VIb is        optionally substituted, on any ring atom capable of        substitution, with 1-3 substituents selected from the group        consisting of alkyl, substituted alkyl, alkoxy, substituted        alkoxy, acyl, acylamino, thiocarbonylamino, acyloxy, amino,        substituted amino, amidino, alkyl amidino, thioamidino,        aminoacyl, aminocarbonylamino, aminothiocarbonylamino,        aminocarbonyloxy, aryl, substituted aryl, aryloxy, substituted        aryloxy, aryloxyaryl, substituted aryloxyaryl, cyano, halogen,        hydroxyl, nitro, oxo, carboxyl, cycloalkyl, substituted        cycloalkyl, guanidino, guanidinosulfone, thiol, thioalkyl,        substituted thioalkyl, thioaryl, substituted thioaryl,        thiocycloalkyl, substituted thiocycloalkyl, thioheteroaryl,        substituted thioheteroaryl, thioheterocyclic, substituted        thioheterocyclic, heteroaryl, substituted heteroaryl,        heterocyclic, substituted heterocyclic, cycloalkoxy, substituted        cycloalkoxy, heteroaryloxy, substituted heteroaryloxy,        heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino,        oxythiocarbonylamino, —OS(O)₂-alkyl, —OS(O)₂-substituted alkyl,        —OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,        —OS(O)₂-substituted heteroaryl, —OS(O)₂-heterocyclic,        —OS(O)₂-substituted heterocyclic, —OSO₂—NRR where each R is        independently hydrogen or alkyl, —NRS(O)₂-alkyl,        —NRS(O)₂-substituted alkyl, —NRS(O)₂-aryl, —NRS(O)₂-substituted        aryl, —NRS(O)₂-heteroaryl, —NRS(O)₂-substituted heteroaryl,        —NRS(O)₂-heterocyclic, —NRS(O)₂-substituted heterocyclic,        —NRS(O)₂—NR-alkyl, —NRS(O)₂—NR-substituted alkyl,        —NRS(O)₂—NR-aryl, —NRS(O)₂—NR-substituted aryl,        —NRS(O)₂—NR-heteroaryl, —NRS(O)₂—NR-substituted heteroaryl,        —NRS(O)₂—NR-heterocyclic, —NRS(O)₂—NR-substituted heterocyclic        where R is hydrogen or alkyl, —N[S(O)₂—R′]₂ and —N[S(O)₂—NR′]₂        where each R′ is independently selected from the group        consisting of alkyl, substituted alkyl, aryl, substituted aryl,        heteroaryl, substituted heteroaryl, heterocyclic and substituted        heterocyclic;    -   R¹³ is selected from the group consisting of hydrogen, C₁₋₁₀        alkyl, Cy, and Cy-C₁₋₁₀ alkyl, wherein alkyl is optionally        substituted with one to four substituents independently selected        from R^(a); and Cy is optionally substituted with one to four        substituents independently selected from R^(b);    -   R¹⁴ is selected from the group consisting of hydrogen, C₁₋₁₀        alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, Cy, Cy-C₁₋₁₀ alkyl,        Cy-C₂₋₁₀ alkenyl and Cy-C₂₋₁₀ alkynyl, wherein alkyl, alkenyl,        and alkynyl are optionally substituted with one to four        substituents selected from phenyl and R^(X), and Cy is        optionally substituted with one to four substituents        independently selected from R^(y);    -   or R¹³, R¹⁴ and the atoms to which they are attached together        form a mono- or bicyclic ring containing 0-2 additional        heteratoms selected from N, O and S;    -   R¹⁵ is selected from the group consisting of C₁₋₁₀ alkyl, C₂₋₁₀        alkenyl, C₂₋₁₀ alkynyl, aryl, aryl-C₁₋₁₀ alkyl, heteroaryl,        heteroaryl-C₁₁₀ alkyl, wherein alkyl, alkenyl and alkynyl are        optionally substituted with one to four substituents selected        from R^(x), and aryl and heteroaryl are optionally substituted        with one to four substituents independently selected from R^(y);    -   or R¹⁴, R¹⁵ and the carbon to which they are attached form a 3-7        membered mono- or bicyclic ring containing 0-2 heteroatoms        selected from N, O and S;    -   R^(a) is selected from the group consisting of Cy and a group        selected from R^(x), wherein Cy is optionally substituted with        one to four substituents independently selected from R^(c;)    -   R^(b) is selected from the group consisting of R^(a), C₁₋₁₀        alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, aryl C₁₋₁₀alkyl, heteroaryl        C₁₋₁₀ alkyl, wherein alkyl, alkenyl, alkynyl, aryl, heteroaryl        are optionally substituted with a group independently selected        from R^(c);    -   R^(c) is selected from the group consisting of halogen, NO₂,        C(O)OR^(f), C₁₋₄ alkyl, C₁₋₄ alkoxy, aryl, aryl C₁₋₄ alkyl,        aryloxy, heteroaryl, NR^(f)R^(g), R^(f)C(O)R^(g),        NR^(f)C(O)NR^(f)R^(g), and CN;    -   R^(d) and R^(e) are independently selected from hydrogen, C₁₋₁₀        alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, Cy and Cy C₁₋₁₀alkyl,        wherein alkyl, alkenyl, alkynyl and Cy are optionally        substituted with one to four substituents independently selected        from R^(c);    -   or R^(d) and R^(e) together with the atoms to which they are        attached form a heterocyclic ring of 5 to 7 members containing        0-2 additional heteroatoms independently selected from oxygen,        sulfur and nitrogen;    -   R^(f) and R^(g) are independently selected from hydrogen, C₁₋₁₀        alkyl, Cy and Cy-C₁₋₁₀ alkyl wherein Cy is optionally        substituted with C₁₋₁₀ alkyl; or R^(f) and R^(g) together with        the carbon to which they are attached form a ring of 5 to 7        members containing 0-2 heteroatoms independently selected from        oxygen, sulfur and nitrogen;    -   R^(h) is selected from the group consisting of hydrogen, C₁₋₁₀        alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, cyano, aryl, aryl C₁₋₁₀        alkyl, heteroaryl, heteroaryl C₁₋₁₀ alkyl, and —SO₂R^(i);        wherein alkyl, alkenyl, and alkynl are optionally substituted        with one to four substitutents independently selected from        R^(a); and aryl and heteroaryl are each optionally substituted        with one to four substituents independently selected from R^(b);    -   R^(i) is selected from the group consisting of C₁₋₁₀ alkyl,        C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, and aryl; wherein alkyl, alkenyl,        alkynyl and aryl are each optionally substituted with one to        four substituents independently selected from R^(c);    -   R^(x) is selected from the group consisting of —OR^(d), —NO₂,        halogen, —S(O)_(m)R^(d), SR^(d), —S(O)₂OR, —S(O)_(m)NR^(d)R^(e),        —NR^(d)R^(e), —O(CR^(f)R^(g))_(n)NR^(d)C(O)R^(d)—CO₂R^(d),        —CO₂(CR^(f)R^(g))_(n)CONR^(d)R^(e), —OC(O)R^(d), —CN,        —C(O)NR^(d)R^(e), —NR^(d)C(O)R^(e), —OC(O)NR^(d)R^(e),        NR^(d)C(O)OR^(e), —NR^(d)C(O)NR^(d)R^(e), —CR^(d)(N—OR^(e)),        CF₃, oxo, NR^(d)C(O)NR^(d)SO₂R^(i), NR^(d)S(O)_(m)R^(e),        —OS(O)₂OR^(d), and —OP(O)(OR^(d))₂;    -   R^(y) is selected from the group consisting of R^(x), C₁₋₁₀        alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, aryl C₁₋₁₀alkyl, heteroaryl        C₁₋₁₀ alkyl, cycloalkyl, heterocyclyl; wherein alkyl, alkenyl,        alkynyl and aryl are each optionally substituted with one to        four substitutents independently selected from R^(x);    -   Cy is cycloalkyl, heterocyclyl, aryl, or heteroaryl;    -   m is an integer from 1 to 2;    -   n is an integer from 1 to 10;    -   W is selected from the group consisting of carbon and nitrogen;    -   W′ is selected from the group consisting of carbon, nitrogen,        oxygen, sulfur, S(O) and S(O)₂;    -   X′ is selected from the group consisting of —C(O)OR^(d),        —P(O)(OR^(d))(OR^(e)), —P(O)(R^(d))(OR^(e)), —S(O)_(m)OR^(d),        —C(O)NR^(d)R^(h), and -5-tetrazolyl;    -   and enantiomers, diastereomers and pharmaceutically acceptable        salts thereof;    -   and further wherein the compound of Formula VIa and/or VIb has a        binding affinity to VLA-4 as expressed by an IC₅₀ of about 15 μM        or less.

Preferably, R¹ and R², together with the carbon atom and W to which theyare bound respectively, ate joined to form a heteroaryl or substitutedheteroaryl group having two nitrogen atoms in the heteroaryl ring.Optionally, the heteroaryl ring may contain other heteroatoms such asoxygen or sulfur. More preferably, R¹ and R², together with the carbonatom and W to which they are bound respectively, are joined to form apyridazine, pyrimidine, pyrazine, 1-oxo-,2,5-thiadiazole or1,1-dioxo-1,2,5-thiadiazole ring; more preferably, a pyrimidine,pyrazine, 1-oxo-1,2,5-thiadiazole or 1,1-dioxo-1,2,5-thiadiazole ring;wherein the pyridazine, pyrimidine, pyrazine, 1-oxo-1,2,5-thiadiazole or1,1-dioxo-1,2,5-thiadiazole ring is optionally substituted with 1 to 3substituents selected from the group consisting of alkyl, substitutedalkyl, alkoxy, substituted alkoxy, amino, substituted amino, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic and halogen.

Preferably, X′ is —C(O)OR^(d).

In another preferred embodiment, the compounds that can be utilized ascombination therapies with methotrexate for the treatment of RA arecompounds defined by Formula VIIa, VIb, VIc, VId, or VIIe:

-   -   wherein R¹³, R¹⁴, R¹⁵ and X′ are as defined herein;    -   R⁵ is selected from the group consisting of alkyl, substituted        alkyl, alkenyl, substituted alkenyl, aryl, substituted aryl,        cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted        cycloalkenyl, heterocyclic, substituted heterocylic, heteroaryl        and substituted heteroaryl;    -   R⁶ is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, cycloalkyl, substituted cycloalkyl,        cycloalkenyl, substituted cycloalkenyl, heterocyclic,        substituted heterocyclic, aryl, substituted aryl, heteroaryl,        substituted heteroaryl, and —SO₂R¹⁰ where R¹⁰ is selected from        the group consisting of alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl,        heterocyclic, substituted heterocyclic, aryl, substituted aryl,        heteroaryl, substituted heteroaryl; and    -   R⁷ and R⁸ are independently selected from the group consisting        of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted        cycloalkyl, amino, substituted amino, aryl, substituted aryl,        heteroaryl, substituted heteroaryl, heterocyclic, substituted        heterocyclic and halogen;    -   R¹⁶ and R¹⁷ are independently selected from the group consisting        of hydrogen, alkyl, substituted alkyl, alkoxy, substituted        alkoxy, amino, substituted amino, cycloalkyl, substituted        cycloalkyl, aryl, substituted aryl, heteroaryl, substituted        heteroaryl, heterocyclic, substituted heterocyclic and halogen;        and    -   R¹⁸ is selected from the group consisting of alkyl, substituted        alkyl, alkoxy, substituted alkoxy, amino, substituted amino,        cycloalkyl, substituted cycloalkyl, aryl, substituted aryl,        heteroaryl, substituted heteroaryl, heterocyclic and substituted        heterocyclic;    -   R²⁰ is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, alkoxy, substituted alkoxy, cycloalkyl,        substituted cycloalkyl, aryl, substituted aryl, heteroaryl,        substituted heteroaryl, heterocyclic, substituted heterocyclic        and halogen;    -   R²¹ is selected from the group consisting of alkyl, substituted        alkyl, alkoxy, substituted alkoxy, amino, substituted amino,        cycloalkyl, substituted cycloalkyl, aryl, substituted aryl,        heterocyclic and substituted heterocyclic;    -   b is 1 or 2;    -   and enatiomers, diastereomers and pharmaceutically acceptable        salts thereof.

Preferably, the compounds that can be utilized as combination therapieswith methotrexate for the treatment of RA are compounds of Formula VIIa,VIIc, or VIId.

In another embodiment, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compoundsdefined by Formula VIIIa and/or VIIIb:

-   -   wherein, in Formula VIIIa, R¹ and R², together with the carbon        atom and W to which they are bound respectively, are joined to        form an aryl, cycloalkenyl, heteroaryl or heterocyclic group        having at least five atoms in the aryl, cycldalkenyl, heteroaryl        or heterocyclic group and optionally containing or additionally        containing in the case of heteroaryl and heterocyclic groups 1        to 3 heteroatoms selected from the group consisting of oxygen,        nitrogen and sulfur, and wherein the heteroaryl or heterocyclic        group is mono-cyclic;    -   in Formula VIIIb, R¹ and R², together with the carbon atom and        W′ to which they are bound respectively, are joined to form a        cycloalkyl, cycloalkenyl or heterocyclic group having at least        five atoms in the cycloalkyl, cycloalkenyl or heterocyclic group        and optionally containing or additionally containing in the case        of the heterocyclic group 1 to 3 heteroatoms selected from the        group consisting of oxygen, nitrogen and sulfur, and wherein the        heterocyclic group is mono-cyclic;    -   and further wherein said aryl, cycloalkyl, cycloalkenyl,        heteroaryl or heterocyclic group of Formula VIIIa or VIIIb is        optionally substituted, on any ring atom capable of        substitution, with 1-3 substituents selected from the group        consisting of alkyl, substituted alkyl, alkoxy, substituted        alkoxy, acyl, acylamino, thiocarbonylamino, acyloxy, amino,        substituted amino, amidino, alkyl amidino, thioamidino,        aminoacyl, aminocarbonylamino, aminothiocarbonylamino,        aminocarbonyloxy, aryl, substituted aryl, aryloxy, substituted        aryloxy, aryloxyaryl, substituted aryloxyaryl, cyano, halogen,        hydroxyl, nitro, oxo, carboxyl, cycloalkyl, substituted        cycloalkyl, guanidino, guanidinosulfone, thiol, thioalkyl,        substituted thioalkyl, thioaryl, substituted thioaryl,        thiocycloalkyl, substituted thiocycloalkyl, thioheteroaryl,        substituted thioheteroaryl, thioheterocyclic, substituted        thioheterocyclic, heteroaryl, substituted heteroaryl,        heterocyclic, substituted heterocyclic, cycloalkoxy, substituted        cycloalkoxy, heteroaryloxy, substituted heteroaryloxy,        heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino,        oxythiocarbonylamino, —OS(O)₂-alkyl, —OS(O)₂-substituted alkyl,        —OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,        —OS(O)₂-substituted heteroaryl, —OS(O)₂-heterocyclic,        —OS(O)₂-substituted heterocyclic, —OSO₂—NRR where each R is        independently hydrogen or alkyl, —NRS(O)₂-alkyl,        —NRS(O)₂-substituted alkyl, —NRS(O)₂-aryl, —NRS(O)₂-substituted        aryl, —NRS(O)₂-heteroaryl, —NRS(O)₂-substituted heteroaryl,        —NRS(O)₂-heterocyclic, —NRS(O)₂-substituted heterocyclic,        —NRS(O)₂—NR-alkyl, —NRS(O)₂—NR-substituted alkyl,        —NRS(O)₂—NR-aryl, —NRS(O)₂—NR-substituted aryl,        —NRS(O)₂—NR-heteroaryl, —NRS(O)₂—NR-substituted heteroaryl,        —NRS(O)₂—NR-heterocyclic, —NRS(O)₂—NR-substituted heterocyclic        where R is hydrogen or alkyl, —N[S(O)₂—R′]₂ and —N[S(O)₂—NR′]₂        where each R′ is independently selected from the group        consisting of alkyl, substituted alkyl, aryl, substituted aryl,        heteroaryl, substituted heteroaryl, heterocyclic and substituted        heterocyclic;    -   R²³ is selected from the group consisting of hydrogen, C₁₋₁₀        alkyl optionally substituted with one to four substituents        independently selected from R^(a′) and Cy optionally substituted        with one to four substituents independently selected from        R^(b′);    -   R²⁴ is selected from the group consisting of Ar¹—Ar²-C₂₋₁₀        alkyl, Ar¹—Ar²-C₂₋₁₀ alkenyl, Ar¹—Ar²-C₂₋₁₀ alkynyl, wherein Ar¹        and Ar² are independently aryl or heteroaryl each of which is        optionally substituted with one to four substituents        independently selected from Rb′; alkyl, alkenyl and alkynyl are        optionally substituted with one to four substituents        independently selected from R^(a);    -   R²⁵ is selected from the group consisting of hydrogen, C₁₋₁₀        alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, aryl, aryl C₁₋₁₀alkyl,        heteroaryl, and heteroaryl C₁₋₁₀ alkyl, wherein alkyl, alkenyl        and alkynyl are optionally substituted with one to four        substituents selected from R^(a′), and aryl and heteroaryl are        optionally substituted with one to four substituents        independently selected from R^(b′);    -   R^(a′) is selected from the group consisting of Cy, —OR^(d′),        —NO₂, halogen —S(O)_(m)R^(d), —SR^(d), —S(O)₂OR^(d′),        —S(O)_(m)NR^(d′)R^(e′), —NR^(d′)R^(e′),        —O(CR^(f′)R^(g′))NR^(d′)R^(e′), —C(O)R^(d′),        —CO₂R^(d′)—CO₂(CR^(f′)R^(g′))_(n)CONR^(d′)R^(e′)—OC(O)R^(d′),        —CN, —C(O)NR^(d′)R^(e′), —NR^(d′)C(O)R^(e′),        —OC(O)NR^(d′)R^(e′)—, NR^(d′)C(O)OR^(e′)—NR^(d′)        C(O)NR^(d′)R^(e′)—CR^(d′)(OR^(e′)), CF₃, and —OCF₃;    -   wherein Cy is optionally substituted with one to four        substituents independently selected from R^(c′);    -   R^(b′) is selected from the group consisting of R^(a′), C₁₋₁₀        alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, aryl C₁₋₁₀ alkyl,        heteroaryl C₁₋₁₀ alkyl,    -   wherein alkyl, alkenyl, aryl, heteroaryl are optionally        substituted with a group independently selected from R^(c′);    -   R^(c′) is selected from the group consisting of halogen, amino,        carboxy, C₁₋₄ alkyl, C₁₋₄ alkoxy, aryl, aryl C₁₋₄ alkyl,        hydroxy, CF₃, and aryloxy;    -   R^(d′) and R^(e′) are independently selected from hydrogen,        C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, Cy and Cy C₁₋₁₀        alkyl, wherein alkyl, alkenyl, alkynyl and Cy are optionally        substituted with one to four substituents independently selected        from R_(c′); or R^(d′) and R^(e′) together with the atoms to        which they are attached form a heterocyclic ring of 5 to 7        members containing 0-2 additional heteroatoms independently        selected from oxygen, sulfur and nitrogen;    -   R^(f′) and R^(g′) are independently selected from hydrogen,        C₁₋₁₀ alkyl, Cy and Cy-C₁₋₁₀ alkyl; or R^(f′) and R^(g′)        together with the carbon to which they are attached form a ring        of 5 to 7 members containing 0-2 heteroatoms independently        selected from oxygen, sulfur and nitrogen;    -   R^(h′) is selected from the group consisting of hydrogen, C₁₋₁₀        alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, cyano, aryl, aryl C₁₋₁₀        alkyl, heteroaryl, heteroaryl C₁₋₁₀ alkyl, or —SO₂R^(i′);    -   wherein alkyl, alkenyl, and alkynyl are optionally substituted        with one to four substitutents independently selected from        R^(a′); and aryl and heteroaryl are each optionally substituted        with one to four substituents independently selected from        R^(b′);    -   R^(i′) is selected from the group consisting of C₁₋₁₀ alkyl,        C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, and aryl;    -   wherein alkyl, alkenyl, alkynyl and aryl are each optionally        substituted with one to four substituents independently selected        from R^(c′);    -   Cy is cycloalkyl, heterocyclyl, aryl, or heteroaryl;    -   X″ is selected from the group consisting of —C(O)OR^(d′),        —P(O)(OR^(d′))(OR^(e′)), —P(O)(R^(d′))(OR^(e′)),        —S(O)_(m)OR^(d′), —C(O)NR^(d′)R^(h′), and -5-tetrazolyl;    -   m is an integer from 1 to 2;    -   n is an integer from 1 to 10;    -   and enantiomers, diastereomers and pharmaceutically acceptable        salts thereof;    -   and further wherein the compound of Formula VIIIa and/or VIIIb        has a binding affinity to VLA-4 as expressed by an IC₅₀ of about        15 μM or less.

Preferably, R¹ and R², together with the carbon atom and W to which theyare bound respectively, are joined to form a heteroaryl or substitutedheteroaryl group having two nitrogen atoms in the heteroaryl ring.Optionally, the heteroaryl ring may contain other heteroatoms such asoxygen or sulfur. More preferably, R¹ and R², together with the carbonatom and W to which they are bound respectively, are joined to form apyridazine, pyrimidine, pyrazine, 1-oxo-1,2,5-thiadiazole or1,1-dioxo-1,2,5-thiadiazole ring; more preferably, a pyrimidine,pyrazine, 1-oxo-1,2,5-thiadiazole or 1,1-dioxo-1,2,5-thiadiazole ring;wherein the pyridazine, pyrimidine, pyrazine, 1-oxo-1,2,5-thiadiazole or1,1-dioxo-1,2,5-thiadiazole ring is optionally substituted with 1 to 3substituents selected from the group consisting of alkyl, substitutedalk-yl, alkoxy, substituted alkoxy, amino, substituted amino,cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic, substituted heterocyclic andhalogen.

Preferably, X″ is —C(O)OR^(d′).

Preferably, R²⁴ is —CH₂—Ar²—Ar¹ and R²⁵ is hydrogen.

In another preferred embodiment, the compounds that can be utilized ascombination therapies with methotrexate for the treatment of RA arecompounds defined by Formula IXa, IXb, IXc, IXd, or IXe:

-   -   wherein R²³, R²⁴, R²⁵ and X″ are as defined herein;    -   R⁵ is selected from the group consisting of alkyl, substituted        alkyl, alkenyl, substituted alkenyl, aryl, substituted aryl,        cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted        cycloalkenyl, heterocyclic, substituted heterocylic, heteroaryl        and substituted heteroaryl;    -   R⁶ is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, cycloalkyl, substituted cycloalkyl,        cycloalkenyl, substituted cycloalkenyl, heterocyclic,        substituted heterocyclic, aryl, substituted aryl, heteroaryl,        substituted heteroaryl, and —SO₂R¹⁰ where R¹⁰ is selected from        the group consisting of alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl,        heterocyclic, substituted heterocyclic, aryl, substituted aryl,        heteroaryl, substituted heteroaryl; and    -   R⁷ and R⁸ are independently selected from the group consisting        of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted        cycloalkyl, amino, substituted amino, aryl, substituted aryl,        heteroaryl, substituted heteroaryl, heterocyclic, substituted        heterocyclic and halogen;    -   R¹⁶ and R¹⁷ are independently selected from the group consisting        of hydrogen, alkyl, substituted alkyl, alkoxy, substituted        alkoxy, amino, substituted amino, cycloalkyl, substituted        cycloalkyl, aryl, substituted aryl, heteroaryl, substituted        heteroaryl, heterocyclic, substituted heterocyclic and halogen;        and    -   R¹⁸ is selected from the group consisting of alkyl, substituted        alkyl, alkoxy, substituted alkoxy, amino, substituted amino,        cycloalkyl, substituted cycloalkyl, aryl, substituted aryl,        heteroaryl, substituted heteroaryl, heterocyclic and substituted        heterocyclic;    -   R²⁰ is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, alkoxy, substituted alkoxy, cycloalkyl,        substituted cycloalkyl, aryl, substituted aryl, heteroaryl,        substituted heteroaryl, heterocyclic, substituted heterocyclic        and halogen;    -   R²¹ is selected from the group consisting of alkyl, substituted        alkyl, alkoxy, substituted alkoxy, amino, substituted amino,        cycloalkyl, substituted cycloalkyl, aryl, substituted aryl,        heterocyclic and substituted heterocyclic;    -   b is 1 or 2;    -   and enantiomers, diastereomers and pharmaceutically acceptable        salts thereof.

Preferably, the compounds that can be utilized as combination therapieswith methotrexate for the treatment of RA are compounds of Formula IXa,IXc, or IXd.

In the above compounds, when X is other than —OH or pharmaceutical saltsthereof, X is preferably a substituent which will convert (e.g.,hydrolyze, metabolize, etc.) in vivo to a compound where X is —OH or asalt thereof. Accordingly, suitable X groups are any art recognizedpharmaceutically acceptable groups which will hydrolyze or otherwiseconvert in vivo to a hydroxyl group or a salt thereof including, by wayof example, esters (X is alkoxy, substituted alkoxy, cycloalkoxy,substituted cycloalkoxy, alkenoxy, substituted alkenoxy, cycloalkenoxy,substituted cycloalkenoxy, aryloxy, substituted aryloxy, heteroaryloxy,substituted heteroaryloxy, heterocyclooxy, substituted heterocyclooxy,and the like).

Unless otherwise defined, R³ and R¹⁵ in the above compounds arepreferably selected from all possible isomers arising by substitutionwith the following groups:

-   -   4-methylbenzyl,    -   4-hydroxybenzyl,    -   4-methoxybenzyl,    -   4-t-butoxybenzyl,    -   4-benzyloxybenzyl,    -   4-[(φ-CH(CH₃)O-]benzyl,    -   4-[(φ-CH(COOH)O-]benzy],    -   4-[BocNHCH₂C(O)NH-]benzyl,    -   4-chlorobenzyl,    -   4-[NH₂CH₂C(O)NH-]benzyl,    -   4-carboxybenzyl,    -   4-[CbzNHCH₂CH₂NH-]benzyl,    -   3-hydroxy-4-((φ-OC(O)NH—)benzyl,    -   4-[HOOCCH₂CH₂C(O)NH-]benzyl, benzyl,    -   4-[2′-carboxylphenoxy-]benzyl,    -   4-[(φ-C(O)NH-]benzyl,    -   3-carboxybenzyl,    -   4-iodobenzyl,    -   4-hydroxy-3,5-diiodobenzyl,    -   4-hydroxy-3-iodobenzyl,    -   4-[2′-carboxyphenyl-]benzyl,    -   φ-CH₂CH₂—,    -   4-nitrobenzyl,    -   2-carboxybenzyl,    -   4-[dibenzylamino]-benzyl,    -   4-[(1′-cyclopropylpiperidin-4′-yl)C(O)NH-]benzyl,    -   4-[—NHC(O)CH₂NHBoc]benzyl,    -   4-carboxybenzyl,    -   4-hydroxy-3-nitrobenzyl,    -   4-[—NHC(O)CH(CH₃)NHBoc]benzyl,    -   4-[—NHC(O)CH(CH₂φ)NHBoc]benzyl,    -   isobutyl,    -   methyl,    -   4—[CH₃C(O)NH-]benzyl,    -   —CH₂-(3-indolyl),    -   n-butyl,    -   t-butyl-OC(O)CH₂—,    -   t-butyl-OC(O)CH₂CH₂—,    -   H₂NC(O)CH₂—,    -   H₂NC(O)CH₂CH₂—,    -   BocNH—(CH₂)₄—,    -   t-butyl-OC(O)—(CH₂)₂—,    -   HOOCCH₂—,    -   HOOC(CH₂)₂—,    -   H₂N(CH₂)₄—,    -   isopropyl,    -   (1-naphthyl)-CH₂—,    -   (2-naphthyl)-CH₂—,    -   (2-thiophenyl)-CH₂—,    -   (φ-CH₂—OC(O)NH—(CH₂)₄—,    -   cyclohexyl-CH₂—,    -   benzyloxy-CH₂—,    -   HOCH₂—,    -   5-(3-N-benzyl)imidazolyl-CH₂—,    -   2-pyridyl-CH₂—,    -   3-pyridyl-CH₂—,    -   4-pyridyl-CH₂—,    -   5-(3-N-methyl)imidazolyl-CH₂—,    -   N-benzylpiperid-4-yl-CH₂—,    -   N-Boc-piperidin-4-yl-CH₂—,    -   N-(phenyl-carbonyl)piperidin-4-yl-CH₂—,    -   H₃CSCH₂CH₂—,    -   1-N-benzylimidazol-4-yl-CH₂—,    -   iso-propyl-C(O)NH—(CH₂)₄—,    -   iso-butyl-C(O)NH—(CH₂)₄—,    -   phenyl-C(O)NH—(CH₂)₄—,    -   benzyl-C(O)NH—(CH₂)₄—,    -   allyl-C(O)NH—(CH₂)₄—,    -   4-(3-N-methylimidazolyl)-CH₂—,    -   4-imidazolyl,    -   4-[(CH₃)₂NCH₂CH₂CH₂—O-]benzyl,    -   4-[(benzyl)₂N-]-benzyl,    -   4-aminobenzyl,    -   allyloxy-C(O)NH(CH₂)₄—,    -   allyloxy-C(O)NH(CH₂)₃—,    -   allyloxy-C(O)NH(CH₂)₂—,    -   NH₂C(O)CH₂—,    -   φ-CH═,    -   2-pyridyl-C(O)NH—(CH₂)₄—,    -   4-methylpyrid-3-yl-C(O)NH—(CH₂)₄—,    -   3-methylthien-2-yl-C(O)NH—(CH₂)₄—,    -   2-pyrrolyl-C(O)NH—(CH₂)₄—,    -   2-furanyl-C(O)NH—(CH₂)₄—,    -   4-methylphenyl—SO₂—N(CH₃)CH₂C(O)NH(CH₂)₄—,    -   4-[cyclopentylacetylenyl]-benzyl,    -   4-[-NHC(O)—(N-Boc)-pyrrolidin-2-yl)]-benzyl-,    -   1-N-methylimidazol-4-yl-CH₂—,    -   1-N-methylimidazol-5-yl-CH₂—,    -   imidazol-5-yl-CH₂—,    -   6-methylpyrid-3-yl-C(O)NH—(CH₂)₄—,    -   4-[2′-carboxymethylphenyl]-benzyl,    -   4-[—NHC(O)NHCH₂CH₂CH₂-φ]-benzyl,    -   4-[-NHC(O)NHCH₂CH₂-(φ]-benzyl,    -   —CH₂C(O)NH(CH₂)₄φ,    -   4-[(φ(CH₂)₄φ]-benzyl,    -   4-[—C≡C-(φ-4′(φ]-benzyl,    -   4-[—C≡C—CH₂—O—S(O)₂-4′-CH₃-φ]-benzyl,    -   4-[—C≡C—CH₂NHC(O)NH₂]-benzyl,    -   4-[—C≡C—CH₂—O-4′-COOCH₂CH₃-(φ]-benzyl,    -   4-[—C≡C—CH(NH₂)-cyclohexyl]-benzyl,    -   —(CH₂)₄NHC(O)CH₂-3-indolyl,    -   —(CH₂)₄NHC(O)CH₂CH₂-3-indolyl,    -   —(CH₂)₄NHC(O)-3-(5-methoxyindolyl),    -   —(CH₂)₄NHC(O)-3-(1-methylindolyl),    -   —(CH₂)₄NHC(O)-4-(—SO₂(CH₃)-φ),    -   —(CH₂)₄NHC(O)-4-(C(O)CH₃)-phenyl,    -   —(CH₂)₄NHC(O)-4-fluorophenyl,    -   —(CH₂)₄NHC(O)CH₂O-4-fluorophenyl,    -   4-[—C≡C-(2-pyridyl)]benzyl,    -   4-[—C≡C—CH₂—O-phenyl]benzyl,    -   4-[—C≡C—CH₂OCH₃]benzyl,    -   4-[—C≡C-(3-hydroxyphenyl)]benzyl,    -   4-[—C≡C—CH₂—O-4′-(—C(O)OC₂H₅)phenyl]benzyl,    -   4-[—C≡C—CH₂CH(C(O)OCH₃)₂]benzyl,    -   4-[—C≡C—CH₂NH—(4,5-dihydro-4-oxo-5-phenyl-oxazo]-2-yl),        3-aminobenzyl,    -   4-[—C≡C—CH₂CH( NHC(O)CH₃)C(O)OH]-benzyl,    -   —H₂C(O)NHCH(CH₃)(φ,    -   —CH₂C(O)NHCH₂-(4-dimethylamino)-(φ,    -   —CH₂C(O)NHCH₂-4-nitrophenyl,    -   —CH₂CH₂C(O)N(CH₃)CH₂—,    -   —CH₂CH₂C(O)NHCH₂CH₂-(N-methyl)-2-pyrrolyl,    -   —CH₂CH₂C(O)NHCH₂CH₂CH₂CH₃,    -   —CH₂CH₂C(O)NHCH₂CH₂-3-indolyl,    -   —CH₂C(O)N(CH₃)CH₂phenyl,    -   —CH₂C(O)NH(CH₂)₂-(N-methyl)-2-pyrrolyl,    -   —CH₂C(O)NHCH₂CH₂CH₂CH₃,    -   —CH₂C(O)NHCH₂CH₂-3-indolyl,    -   —(CH₂)₂C(O)NHCH(CH₃)    -   —(CH₂)₂C(O)NHCH₂-4-dimethylaminophenyl,    -   —(CH₂)₂C(O)NHCH₂-4-nitrophenyl,    -   —CH₂C(O)NH-4-[-NHC(O)CH₃-phenyl],    -   —CH₂C(O)NH-4-pyridyl,    -   —CH₂C(O)NH-4-[dimethylaminophenyl],    -   —CH₂C(O)NH-3-methoxyphenyl,    -   —CH₂CH₂C(O)NH-4-chlorophenyl,    -   —CH₂CH₂C(O)NH-2-pyridyl,    -   —CH₂CH₂C(O)NH-4-methoxyphenyl,    -   —CH₂CH₂C(O)NH-3-pyridyl,    -   4-[(CH₃)₂NCH₂CH₂O-]benzyl,    -   —(CH₂)₃NHC(NH)NH—SO₂-4-methylphenyl,    -   4-[(CH₃)₂NCH₂CH₂O-]benzyl,    -   —(CH₂)₄NHC(O)NHCH₂CH₃,    -   —(CH₂)₄NHC(O)NH-phenyl,    -   —(CH₂)₄NHC(O)NH-4-methoxyphenyl,    -   4-[4′-pyridyl-C(O)NH-]benzyl,    -   4-[3 ′-pyridyl-C(O)NH-]benzyl,    -   4-[—NHC(O)NH-3′-methylphenyl]benzyl,    -   4-[—NHC(O)CH₂NHC(O)NH-3′-methylphenyl]benzyl,    -   4-[—NHC(O)-(2′,3′-dihydroindol-2-yl)]benzyl,    -   4-[—NHC(O)-(2′,3′-dihydro-N-Boc-indol-2-yl)]benzyl,    -   4-[—OCH₂CH₂-1′-(4′-pyrimidinyl)-piperazinyl]benzyl,    -   4-[—OCH₂CH₂-(1′-piperidinyl)benzyl,    -   4-[—OCH₂CH₂-(1′-pyrrolidinyl)]benzyl,    -   4-[—OCH₂CH₂CH₂-(1′-piperidinyl)]benzyl-,    -   —CH₂-3-(1,2,4-triazolyl),    -   4-[-OCH₂CH₂CH₂-4-(3′-chlorophenyl)-piperazin-1-yl]benzyl,    -   4-[-OCH₂CH₂N(φ)CH₂CH₃]benzyl,    -   4-[-OCH₂-3′-(N-Boc)-piperidinyl]benzyl,    -   4-[di-n-pentylamino]benzyl,    -   4-[n-pentylamino]benzyl,    -   4-[di-iso-propylamino-CH₂CH₂O-]benzyl,    -   4-[—OCH₂CH₂-(N-prorpholnyl)]benzyl,    -   4-[—O-(3′-(N-Boc)-piperidinyl]benzyl,    -   4-[—OCH₂CH(NHBoc)CH₂cyclohexyl]benzyl,    -   p-[OCH₂CH₂-(N-piperidinyl]benzyl,    -   4-[—OCH₂CH₂CH₂-(4-m-chlorophenyl)-piperazin-1-yl]benzyl,    -   4-[—OCH₂CH₂-(N-homopiperidinyl)benzyl,    -   4-[—NHC(O)-3′-(N-Boc)-piperidinyl]benzyl,    -   4-[—OCH₂CH₂N(benzyl)₂]benzyl,    -   —CH₂-2-thiazolyl,    -   3-hydroxybenzyl,    -   4-[—OCH₂CH₂CH₂N(CH₃)₂]benzyl,    -   4-[—NHC(S)NiCH₂CH₂-(N-morpholino)]benzyl,    -   4-[—OCH₂CH₂N(C₂H₅)₂]benzyl,    -   4-[—OCH₂CH₂CH₂N(C₂H₅)₂]benzyl,    -   4-[CH₃(CH₂)₄NH-]benzyl,    -   4-[N-n-butyl,N-n-pentylamino-]benzyl,    -   4-[—NHC(O)-4′-piperidinyl]benzyl,    -   4-[—NHC(O)CH(NHBoc)(CH₂)₄NHCbz]benzyl,    -   4-[—NHC(O)-(1′,2′,3′,4′-tetrahydro-N-Boc-isoquinolin-1′-yl]benzyl,    -   p-[-OCH₂CH₂CH₂-1′-(4′-methyl)-piperazinyl]benzyl,    -   —(CH₂)₄NH-Boe,    -   3-[—OCH₂CH₂CH₂N(CH₃)₂]benzyl,    -   4-[—OCH₂CH₂CH₂N(CH₃)₂]benzyl,    -   3-[—OCH₂CH₂-(1′-pyrrolidinyl)]benzyl,    -   4-[—OCH₂CH₂CH₂N(CH₃)benzyl]benzyl,    -   4-[—NHC(S)NHCH₂CH₂CH₂-(N-morpholino)]benzyl,    -   4-[—OCH₂CH₂-(N-morpholino)]benzyl,    -   4-[—NHCH₂-(4′-chlorophenyl)]benzyl,    -   4-[—NHC(O)NH-(4′-cyanophenyl)]benzyl,    -   4-[—OCH₂COOH]benzyl,    -   4-[—OCH₂COO-t-butyl]benzyl,    -   4-[—NHC(O)-5′-fluoroindol-2-yl]benzyl,    -   4[-[—NHC(S)NH(CH₂)₂-1-piperidinyl]benzyl,    -   4-[—N(SO₂CH₃)(CH₂)₃—N(CH₃)₂]benzyl,    -   4-[—NHC(O)CH₂CH(C(O)OCH₂(p)-NHCbz]benzyl,    -   4-[—NHS(O)₂CF₃]benzyl,    -   3-[—O-(N-methylpiperidin-4′-yl]benzyl,    -   4-[—C(═NH)NH₂]benzyl,    -   4-[—NHSO₂—CH₂C]benzyl,    -   4-[—NHC(O)-(1′,2′,3′,4′-tetrahydroisoquinolin-2′-yl]benzyl,    -   4-[—NHC(S)NH(CH₂)₃-N-morpholino]benzyl,    -   4-[—NHC(O)CH(CH₂CH₂CH₂CH₂NH₂)NHBoc]benzyl,    -   4-[—C(O)NH₂]benzyl,    -   4-[—NHC(O)NH-3′-methoxyphenyl]benzyl,    -   4-[—OCH₂CH₂-indol-3′-yl]benzyl,    -   4-[—OCH₂C(O)NH-benzyl]benzyl,    -   4-[—OCH₂C(O)O-benzyl]benzyl,    -   4-[—OCH₂C(O)OH]benzyl,    -   4-[—OCH₂-2′-(4′,5′-dihydro)imidazolyl]benzyl,    -   —CH₂C(O)NHCH₂-(4-dimethylamino)phenyl,    -   —CH₂C(O)NHCH₂-(4-dimethylamino)phenyl,    -   4-[—NHC(O)-L-2′-pyrrolidinyl-N—SO₂-4′-methylphenyl]benzyl,    -   4-[—NHC(O)NHCH₂CH₂CH₃]benzyl,    -   4-aminobenzyl]benzyl,    -   4-[—OCH₂CH₂-1-(4-hydroxy-4-(3-methoxypyrrol-2-yl)-piperazinyl]benzyl,    -   4-[—O-(N-methylpiperidin-4′-yl)]benzyl,    -   3-methoxybenzyl,    -   4-[—NHC(O)-piperidin-3′-yl]benzyl,    -   4-[—NHC(O)-pyridin-2′-yl]benzyl,    -   4-[—NHCH₂-(4′-chlorophenyl)]benzyl,    -   4-[—NHC(O)-(N-(4′-CH₃-(p-SO₂)-L-pyrrolidin-2′-yl)]benzyl,    -   4-[-NHC(O)NHCH₂CH₂-(p]benzyl,    -   4-[—OCH₂C(O)NH₂]benzyl,    -   4-[—OCH₂C(O)NH-t-butyl]benzyl,    -   4-[—OCH₂CH₂-1-(4-hydroxy-4-phenyl)-piperidinyl]benzyl,    -   4-[—NHSO₂—CH═CH₂]benzyl,    -   4-[—NHSO₂—CH₂CH₂C]benzyl,    -   —CH₂C(O)NHCH₂CH₂N(CH₃)₂,    -   4-[(1′-Cbz-piperidin-4′-yl)C(O)NH-]benzyl,    -   4-[(1′-Boc-piperidin-4′-yl)C(O)NH-]benzyl,    -   4-[(2′-bromophenyl)C(O)NH-]benzyl,    -   4-[—NHC(O)-pyridin-4′-yl]benzyl,    -   4-[(4′-(CH₃)₂NC(O)O-)phenyl)-C(O)NH-]benzyl,    -   4-[—NHC(O)-1′-methylpiperidin-4′-yl-]benzyl,    -   4-(dimethylamino)benzyl,    -   4-[-NHC(O)-(1′-N-Boc)-piperidin-2′-yl]benzyl,    -   3-[-NHC(O)-pyridin-4′-yl]benzyl,    -   4-[(tert-butyl-O(O)CCH₂—O-benzyl)-NH-]benzyl,    -   [BocNHCH₂C(O)NH-]butyl,    -   4-benzylbenzyl,    -   2-hydroxyethyl,    -   4-[(Et)₂NCH₂CH₂CH₂NHC(S)NH-]benzyl,    -   4-[(1′-Boc-4′-hydroxypyrrolidin-2′-yl)C(O)NH-]benzyl,    -   4-[(pCH₂CH₂CH₂NHC(S)NH-]benzyl,    -   4-[(perhydroindolin-2′-yl)C(O)NH-]benzyl,    -   2-[4-hydroxy-4-(3-methoxythien-2-yl)piperidin-1-yl]ethyl,    -   4-[(1′-Boc-perhydroindolin-2′-yl)-C(O)NH-]benzyl,    -   4-[N-3 -methylbutyl-N-trifluoromethanesulfonyl)amino]benzyl,    -   4-[N-vinylsulfonyl)amino]benzyl,    -   4-[2-(2-azabicyclo[3.2.2]octan-2-yl)ethyl-O-]benzyl,    -   4-[4′-hydroxypyrrolidin-2′-yl)C(O)NH-]benzyl,    -   4-((pNHC(S)NH)benzyl,    -   4-(EtNHC(S)NH)benzyl,    -   4-((pCH₂NHC(S)NH)benzyl,    -   3 -[(1′-Boc-piperidin-2′-yl)C(O)NH-]benzyl,    -   3-[piperidin-2′-yl-C(O)NH-]benzyl,    -   4-[(3′-Boc-thiazolidin-4′-yl)C(O)NH-]benzyl,    -   4-(pyridin-3 ′-yl-NHC(S)NH)benzyl,    -   4-(CH₃-NHC(S)NH)benzyl,    -   4-(H₂NCH₂CH₂CH₂C(O)NH)benzyl,    -   4-(BocHNCH₂CH₂CH₂C(O)NH)benzyl,    -   4-(pyridin-4′-yl-CH₂NH)benzyl,    -   4-[(N,N-di(4-N,N-dimethylamino)benzyl)amino]benzyl,    -   4-[(1-Cbz-piperidin-4-yl)C(O)NH-]butyl,    -   4-[(pCH₂0CH₂(BocHN)CHC(O)NH]benzyl,    -   4-[(piperidin-4′-yl)C(O)NH-]benzyl,    -   4-[(pyrrolidin-2′-yl)C(O)NH-]benzyl,    -   4-(pyridin-3 ′-yl-C(O)NH)butyl,    -   4-(pyridin-4′-yl-C(O)NH)butyl,    -   4-(pyridin-3 ′-yl-C(O)NH)benzyl,    -   4-[CH₃NHCH₂CH₂CH₂C(O)NH-]benzyl,    -   4-[CH₃N(Boc)CH₂CH₂CH₂C(O)NH-]benzyl,    -   4-(aminomethyl)benzyl,    -   4-[(pCH₂OCH₂(H₂N)CHC(O)NH]benzyl,    -   4-[(1′,4′-di(Boc)piperazin-2′-yl)-C(O)NH-]benzyl,    -   4-[(piperazin-2′-yl)-C(O)NH-]benzyl,    -   4-[(N-toluenesulfonylpyrrolidin-2′-yl)C(O)NH-]butyl,    -   4-[—NHC(O)-4′-piperidinyl]butyl,    -   4-[—NHC(O)-1′-N-Boc-piperidin-2′-yl]benzyl,    -   4-[—NHC(O)-piperidin-2′-yl]benzyl,    -   4-[(1′-N-Boc-2′,3′-dihydroindolin-2′-yl)-C(O)NH]benzyl,    -   4-(pyridin-3′-yl-CH₂NH)benzyl,    -   4-[(piperidin-1′-yl)C(O)CH₂—O-]benzyl,    -   4-[(CH₃)₂CH)₂NC(O)CH₂—O-]benzyl,    -   4-[HO(O)C(Cbz-NH)CHCH₂CH₂—C(O)NH-]benzyl,    -   4-[(pCH₂O(O)C(Cbz-NH)CHCH₂CH₂—C(O)NH-]benzyl,    -   4-[-NHC(O)-2′-methoxyphenyl]benzyl,    -   4-[(pyrazin-2′-yl)C(O)NH-]benzyl,    -   4-[HO(O)C(NH₂Y)CHCH₂CH₂—C(O)NH-]benzyl-1l        -(2′-formyl-1′,2′,3′,4′-tetrahydroisoquinolin-3′-yl-CH₂NH—)benzyl,    -   N-Cbz-NHCH₂—,    -   4-[(4′-methylpiperazin-1′-yl)C(O)O-]benzyl,    -   4-[CH₃(N-Boc)NCH₂C(O)NH-]benzyl,    -   4-[—NHC(O)-(1′,2′,3′,4′-tetrahydro-N-Boc-isoquinolin-3′-yl]-benzyl,    -   4-[CH₃NHCH₂C(O)NH-]benzyl,    -   (CH₃)₂NC(O)CH₂—,    -   4-(N-methylacetamido)benzyl,    -   4-(1′,2′,3′,4′-tetrahydroisoquinolin-3′-yl-CH₂NH-)benzyl,    -   4-[(CH₃)₂NHCH₂C(O)NH-]benzyl,    -   (1-toluenesulfonylimidizol-4-yl)methyl,    -   4-[(1′-Boc-piperidin-4′-yl)C(O)NH-]benzyl,    -   4-trifluoromethylbenzyl,    -   4-[(2′-bromophenyl)C(O)NH-]benzyl,    -   4-[(CH₃)₂NC(O)NH-]benzyl,    -   4-[CH₃OC(O)NH-]benzyl,    -   4-[(CH₃)₂NC(O)O-]benzyl,    -   4-[(CH₃)₂NC(O)N(CH₃)-]benzyl,    -   4-[CH₃OC(O)N(CH₃)-]benzyl,    -   4-(N-methyltrifluoroacetamido)benzyl,    -   4-[(1′-methoxycarbonylpiperidin-4′-yl)C(O)NH-]benzyl,    -   4-[(4′-phenylpiperidin-4′-yl)C(O)NH-]benzyl,    -   4-[(4′-phenyl-1′-Boc-piperidin-4′-yl)-C(O)NH-]benzyl,    -   4-[(piperidin-4′-yl)C(O)O-]benzyl,    -   4-[(1′-methylpiperidin-4′-yl)-O-]benzyl,    -   4-[( 1′-methylpiperidin-4′-yl)C(O)O-]benzyl,    -   4-[(4′-methylpiperazin- 1′-yl)C(O)NH-]benzyl,    -   3-[(CH₃)₂NC(O)O-]benzyl,    -   4-[(4′-phenyl-1′-Boc-piperidin-4′-yl)-C(O)O-]benzyl,    -   4-(N-toluenesulfonylamino)benzyl,    -   4-[(CH₃)₃CC(O)NH-]benzyl,    -   4-[(morpholin-4′-yl)C(O)NH-]benzyl,    -   4-[(CH₃CH₂)₂NC(O)NH-]benzyl,    -   4-[-C(O)NH-(4′-piperidinyl)]benzyl,    -   4-[(2′-trifluoromethylphenyl)C(O)NH-]benzyl,    -   4-[(2′-methylphenyl)C(O)NH-]benzyl,    -   4-[(CH₃)₂NS(O)₂O-]benzyl,    -   4-[(pyrrolidin-2′-yl)C(O)NH-]benzyl,    -   4-[-NHC(O)-piperidin- 1′-yl]benzyl,    -   4-[(thiomorpholin-4′-yl)C(O)NH-]benzyl,    -   4-[(thiomorpholin-4′-yl sulfone)-C(O)NH-]benzyl,    -   4-[(morpholin-4′-yl)C(O)O-]benzyl,    -   3-nitro-4-(CH₃OC(O)CH₂O-)benzyl,    -   (2-benzoxazolinon-6-yl)methyl-,    -   (2-1,4-benzoxazin-3(4H)-one-7-yl)methyl-,    -   4-[(CH₃)₂NS(O)₂NH-]benzyl,    -   4-[(CH₃)₂NS(O)₂N(CH₃)-]benzyl,    -   4-[(thiomorpholin-4′-yl)C(O)O-]benzyl,    -   4-[(thiomorpholin-4′-yl sulfone)-C(O)O-]benzyl,    -   4-[(piperidin-1′-yl)C(O)O-]benzyl,    -   4-[(pyrrolidin-1′-yl)C(O)O-]benzyl,    -   4-[(4′-methylpiperazin-1′-yl)C(O)O-]benzyl,    -   4-[(2′-methylpyrrolidin-1′-yl)-,    -   (pyridin-4-yl)methyl-,    -   4-[(piperazin-4′-yl)-C(O)O-]benzyl,    -   4-[(1′-Boc-piperazin-4′-yl)-C(O)O-]benzyl,    -   4-[(4′-acetylpiperazin- 1′-yl)C(O)O-]benzyl,    -   p-[(4′-methanesulfonylpiperazin-1′-yl)-benzyl,    -   3-nitro-4-[(morpholin-4′-yl)-C(O)O-]benzyl,    -   4-{[(CH₃)₂NC(S)]₂N-}benzyl,    -   N-Boc-2-aminoethyl-,    -   4-[(1,1-dioxothiomorpholin-4-yl)-C(O)O-]benzyl,    -   4-[(CH₃)₂NS(O)₂-]benzyl,    -   4-(imidazolid-2′-6ne-1′-yl)benzyl,    -   4-[(piperidin-1′-yl)C(O)O-]benzyl,    -   1 -N-benzyl-imidazol-4-yl-CH₂—,    -   3,4-dioxyethylenebenzyl (i.e., 3,4-ethylenedioxybenzyl),    -   3,4-dioxymethylenebenzyl (i.e., 3,4-methylenedioxybenzyl),    -   4-[-N(SO₂)(CH₃)CH₂CH₂CH₂N(CH₃)₂]benzyl,    -   4-(3′-formylimidazolid-2′-one-1′-yl)benzyl,    -   4-[NHC(O)CH(CH₂CH₂CH₂CH₂NH₂)NHBoc]benzyl,    -   [2′-[4″-4″-(3″′-methoxythien-2″′-yl)piperidin-2″-yl]ethoxy]benzyl,        and    -   p-[(CH₃)₂NCH₂CH₂N(CH₃)C(O)O-]benzyl.

Preferably, R⁵ in the above compounds is selected from the groupconsisting of alkyl, substituted alkyl, aryl, substituted aryl,heterocyclic, substituted heterocylic, heteroaryl and substitutedheteroaryl. Even more preferably R⁵ is selected from the groupconsisting of 4-methylphenyl, methyl, benzyl, n-butyl, n-hexyl,4-chlorophenyl, 1-naphthyl, 2-naphthyl, 4-methoxyphenyl, phenyl,2,4,6-trimethylphenyl, 2-(methoxycarbonyl)phenyl, 2-carboxyphenyl,3,5-dichlorophenyl, 4-trifluoromethylphenyl, 3,4-dichlorophenyl,3,4-dimethoxyphenyl, 4-(CH₃C(O)NH-)phenyl, 4-trifluoromethoxyphenyl,4-cyanophenyl, isopropyl, 3,5-di-(trifluoromethyl)phenyl,4-t-butylphenyl, 4-t-butoxyphenyl, 4-nitrophenyl, 2-thienyl,1-N-methyl-3-methyl-5-chloropyrazol-4-yl, phenethyl,1-N-methylimidazol-4-yl, 4-bromophenyl, 4-amidinophenyl,4-methylamidinophenyl, 4-[CH₃SC(═NH)]phenyl, 5-chloro-2-thienyl,2,5-dichloro-4-thienyl, 1-N-methyl-4-pyrazolyl, 2-thiazolyl,5-methyl-1,3,4-thiadiazol-2-yl, 4-[H₂NC(S)]phenyl, 4-aminophenyl,4-fluorophenyl, 2-fluorophenyl, 3-fluorophenyl, 3,5-difluorophenyl,pyridin-3-yl, pyrimidin-2-yl, 4-(3′-dimethylamino-n-propoxy)-phenyl, and1-methylpyrazol-4-yl.

Preferably, R¹³ in the above compounds is selected from hydrogen or C₁₋₆alkyl; more preferably, hydrogen or C₁₋₃ alkyl; and still morepreferably, hydrogen or methyl.

In a preferred embodiment, R¹⁴ in the above compounds is preferablyhydrogen and R¹⁵ is preferably C₁₋₁₀ alkyl or Cy-C₁₋₁₀ alkyl, whereinalkyl is optionally substituted with one to four substituents selectedfrom phenyl and R^(x), and Cy is optionally substituted with one to foursubstituents independently selected from R^(y), or R¹⁴ and R¹⁵ and thecarbon to which they are attached together from a 3-7 membered mono- orbicyclic carbon only ring. For the purpose of R¹⁵, Cy is preferablyaryl, more preferably phenyl. In a preferred embodiment, R¹⁵ isphenyl-C₁₋₃ alkyl, wherein phenyl is optionally substituted with one ortwo groups selected from R^(y). Additional preferred embodiments for R¹⁴and R¹⁵ are disclosed in International Patent Application PublicationNo. WO 98/53814, which application is incorporated herein by referencein its entirety.

In a preferred embodiment of the above compounds, R¹⁶ is substitutedamino; R¹⁷ and/or R²⁰ are hydrogen; and R¹⁸ and/or R²¹ are alkyl,substituted alkyl, aryl or substituted aryl.

In a preferred embodiment, R²³ in the above compounds is hydrogen.Preferably, R²⁴ in the above compounds is Ar¹—Ar²-C₁₋₁₀ alkyl whereinAr¹ and Ar² are optionally substituted with from 1to 4 groupsindependently selected from R^(b) and R²⁵ is hydrogen. More preferably,R²⁴ is Ar¹—Ar²-C₁₋₃ alkyl wherein Arl and Ar2are optionally substitutedwith from 1 to 4 groups independently selected from R^(b); still morepreferably, R²⁴ is —CH₂—Ar²—Ar¹ and R²⁵ is hydrogen. Additionalpreferred embodiments are disclosed in International Patent ApplicationPublication No. WO 98/53817, which application is incorporated herein byreference in its entirety.

Preferably, R³ and R^(3′), or R¹⁴ and R¹⁵, or R²⁴ and R²⁵ are derivedfrom L-amino acids or other similarly configured starting materials.Alternatively, racemic mixtures can be used.

Preferably, x in the above compounds is an integer from 1to 4.

Preferred compounds include those set forth in Tables 3-6 below: TABLE 3

R⁵ R⁶ R⁷ R⁸ R⁹ X 4-CH₃—Ph— H— H— H— 4-(CH₃)₂NC(O)O— —OC(CH₃)₃ 4-CH₃—Ph—H— H— H— 4-(CH₃)₂NC(O)O— —OH 4-CH₃—Ph— CH₃— H— H— 4-(CH₃)₂NC(O)O——OC(CH₃)₃ 4-CH₃—Ph— CH₃— H— H— 4-(CH₃)₂NC(O)O— —OH 4-CH₃—Ph— 4-CH₃—Ph—H— H— 4-(CH₃)₂NC(O)O— —OH 1-CH₃— CH₃— H— H— 4-(CH₃)₂NC(O)O— —OHpyrazol-4-yl- 4-CH₃—Ph— CH₃— H— H— 4-(CH₃)₂NC(O)O— —OCH(CH₃)₂ 3-pyridyl-CH₃— H— H— 4-(CH₃)₂NC(O)O— —OC(CH₃)₃ 1-(n-C₄H₉)— CH₃ H— H—4-(CH₃)₂NC(O)O— —OC(CH₃)₃ pyrazol-4-yl- 4-CH₃—Ph— CH₃ H— H— H— —OH1-(n-C₄H₉)— CH₃— H— H— 4-(CH₃)₂NC(O)O— —OH pyrazol-4-yl- 3-pyridyl- CH₃H— H— 4-(CH₃)₂NC(O)O— —OH 4-CH₃—Ph— CH₃ (CH₃)₂N— H— H— —OH 1-CH₃— CH₃—H— H— 4-(CH₃)₂NC(O)O— —OCH(CH₃)₂ pyrazol-4-yl- 3-pyridyl- CH₃— H— H—4-(1-CH₃-piperazin- —OCH(CH₃)₂ 4-yl)C(O)O— 3-pyridyl- CH₃— H— H—4-(1-CH₃-piperazin- —OC(CH₃)₃ 4-yl)C(O)O— 3-pyridyl- CH₃ H— H—4-(1-CH₃-piperazin- —OH 4-yl)-C(O)O—Ph = phenyl

TABLE 4

R^(16′) R^(20′) R^(18′) R¹⁹ X Cl— H— NO₂— 4-(CH₃)₂NC(O)O— —OH H— H—PhCH₂O— H— —OH H— H— PhCH₂O— 4-(CH₃)₂NC(O)O— —OH H— H— Ph—4-(CH₃)₂NC(O)O— —OH H— H— 3-NO₂—Ph— 4-(CH₃)₂NC(O)O— —OH H— H— 3-pyridyl-4-(CH₃)₂NC(O)O— —OH H— H— 2-PhCH₂CH₂— 4-(CH₃)₂NC(O)O— —OH H— H—2-CH₃—Ph— 4-(CH₃)₂NC(O)O— —OH H— H— (CH₃)₂NC(O)— 4-(CH₃)₂NC(O)O— —OH(CH₂)₂— H— Ph— H— 4-(CH₃)₂NC(O)O— —OH H— 2- H— 4-(CH₃)₂NC(O)O— —OH CF₃—Ph— H— 2- H— 4-(CH₃)₂NC(O)O— —OH HO CH₂ Ph— H— H— CF₃CH₂—4-(CH₃)₂NC(O)O— —OH H— H— PhCH₂— 4-(CH₃)₂NC(O)O— —OH H— H— 2-CH₃—Ph—4-(CH₃)₂NC(O)O— —OCH(CH₃)₂ H— H— 2-PhCH₂CH₂— 4-(CH₃)₂NC(O)O— —OCH(CH₃)₂H— H— 2-PhCH₂CH₂— H— —OCH(CH₃)₂ cyclohexyl- H— H— 4-(CH₃)₂NC(O)O— —OH(CH₃)N— H— H— CH₃CH₂CH₂— 4-(CH₃)₂NC(O)O— —OH H— H— 2-CH₃O—Ph—4-(CH₃)₂NC(O)O— —OH H— H— 2-F—Ph— 4-(CH₃)₂NC(O)O— —OH (CH₃)₂CH— H—2-CH₃—Ph— 4-(CH₃)₂NC(O)O— —OH (CH₃)N— (CH₃)₂CH—NH— H— 2-CH₃—Ph—4-(CH₃)₂NC(O)O— —OH (CH₃)₂CHCH₂— H— 2-CH₃—Ph— 4-(CH₃)₂NC(O)O— —OH(CH₃)N— CH₃CH₂CH₂— H— 2-CH₃—Ph— 4-(CH₃)₂NC(O)O— —OH (CH₃)N— (CH₃)₂N— H—2-CH₃—Ph— 4-(CH₃)₂NC(O)O— —OH cyclohexyl- H— 3-pyridyl- 4-(CH₃)₂NC(O)O——OH (CH₃)N— H— H— 2-PhCF₂CH₂— 4-(CH₃)₂NC(O)O— —OH H— Cl— 2-PhCF₂CH₂—4-(CH₃)₂NC(O)O— —OH (HOCH₂CH₂)₂N— H— H— 4-(CH₃)₂NC(O)O— —OH(HOCH₂CH₂)₂N— H— 2-CH₃—Ph— 4-(CH₃)₂NC(O)O— —OH Ph(CH₃)N— H— 2-CH₃—Ph—4-(CH₃)₂NC(O)O— —OH (CH₃)₂CHO— H— 2-CH₃—Ph— 4-(CH₃)₂NC(O)O— —OH(CH₃)₂CHCH₂— H— 2-CH₃—Ph— 4-(CH₃)₂NC(O)O— —OH CH₂(CH₃)N— CH₃NH— H—2-CH₃—Ph— 4-(CH₃)₂NC(O)O— —OH 2-CH₃—Ph— H— 2-CH₃—Ph— 4-(CH₃)₂NC(O)O— —OHHOCH₂CH₂— H— 2-CH₃—Ph— 4-(CH₃)₂NC(O)O— —OH (CH₃)N— cyclohexyl-NH— H—2-CH₃—Ph— 4-(CH₃)₂NC(O)O— —OH 1-CH₃-piperidin- H— 2-CH₃—Ph—4-(CH₃)₂NC(O)O— —OH 4-yl-(CH₃)N— (CH₃)₂CH— H— 2-CH₃—Ph— 4-(CH₃)₂NC(O)O——OH (CH₃CH₂—)N— H— H— 2,4,6-tri-CH₃—Ph— 4-(CH₃)₂NC(O)O— —OH H— H—(CH₃)₂CH— 4-(CH₃)₂NC(O)O— —OH CH₃(CH₂)₃— H— 2-CH₃—Ph— 4-(CH₃)₂NC(O)O——OH (CH₃)N— CH₃CH₂CH₂— H— 2-CH₃—Ph— 4-(CH₃)₂NC(O)O— —OH (CH₃CH₂—)N—(CH₃CH₂)₂N— H— 2-CH₃—Ph— 4-(CH₃)₂NC(O)O— —OH CH₃CH₂— H— 2-CH₃—Ph—4-(CH₃)₂NC(O)O— —OH (CH₃)N— H— H— cyclohexyl- 4-(CH₃)₂NC(O)O— —OH(furan-2-yl)CH₂— H— 2-CH₃—Ph— 4-(CH₃)₂NC(O)O— —OH (CH₃)N—4-Cl—Ph—(CH₃)N— H— 2-CH₃—Ph— 4-(CH₃)₂NC(O)O— —OH H— H— thien-3-yl-4-(CH₃)₂NC(O)O— —OH H— H— thien-2-yl- 4-(CH₃)₂NC(O)O— —OH HOCH₂CH₂— H—2-F—Ph— 4-(CH₃)₂NC(O)O— —OH (CH₃)N— H— H— piperidin-1-yl-4-(CH₃)₂NC(O)O— —OH H— H— (CH₃CH₂CH₂)₂—CH— 4-(CH₃)₂NC(O)O— —OHcyclobutyl- H— 2-CH₃—Ph— 4-(CH₃)₂NC(O)O— —OH (CH₃)N— H— H— 2-HOCH₂—Ph—4-(CH₃)₂NC(O)O— —OH H— H— 2,6-di-F—Ph— 4-(CH₃)₂NC(O)O— —OH H— H—2,4-di-CH₃O— 4-(CH₃)₂NC(O)O— —OH pyrimidin-5-yl cyclohexyl- H— 2-CH₃—Ph—4-(CH₃)₂NC(O)O— —OH (CH₃)N— H— H— 2-CF₃—Ph— 4-(CH₃)₂NC(O)O— —OHcyclohexyl- H— 2-CH₃O—Ph— 2,6-di-CH₃O—Ph— —OH (CH₃)N— (CH₃)₂CH— H—2-F—Ph— 2,6-di-CH₃O—Ph— —OH (CH₃)N— (CH₃)₂CH— H— 2-F—Ph— 2-CH₃O—Ph— —OH(CH₃)N— cyclohexyl- H— 2,6-di-F—Ph— 2,6-di-F—Ph— —OH (CH₃)N— cyclohexyl-H— 2-HOCH₂—Ph— 2,6-di-CH₃O—Ph— —OH (CH₃)N— (HOCH₂CH₂)₂N— H—2,4,6-tri-CH₃—Ph— 2,6-di-CH₃O—Ph— —OH cyclohexyl- H— 2-CF₃—Ph— 2-NC—Ph——OH (CH₃)N— cyclohexyl- H— thien-3-yl- 2,6-di-CH₃O—Ph— —OH (CH₃)N—cyclohexyl- H— thien-2-yl- 4-CF₃—Ph— —OH (CH₃)N— cyclohexyl- H—3-pyridyl- 2,6-di-CH₃O—Ph— —OH (CH₃)N— cyclohexyl- H— 2-NO₂—Ph—2,6-di-CH₃O—Ph— —OH (CH₃)N— cyclohexyl- H— 2,6-di-Cl—Ph— 2,6-di-CH₃O—Ph——OH (CH₃)N— cyclohexyl- H— 4-pyridyl- 3-HOCH₂—Ph— —OH (CH₃)N— (CH₃)₂CH—H— 2,6-di-CH₃O—Ph— 2,6-di-CH₃O—Ph— —OH (CH₃CH₂—)N— cyclohexyl- H—2,6-di-Cl—Ph— 2,6-di-CH₃O—Ph— —OH (CH₃)N— CH₃CH₂— H— 2,4,6-tri-CH₃—Ph—2-NC—Ph— —OH (CH₃)N— (CH₃)₂CH— H— 2,4,6-tri-CH₃—Ph— 3-pyridyl- —OH(CH₃)N— (HOCH₂CH₂)₂N— H— 2,4,6-tri-CH₃—Ph— 2-NC—Ph— —OH 1-CH₃-piperidin-H— 2-NC—Ph— 2,6-di-F—Ph— —OH 4-yl-(CH₃)N— (CH₃)₂CH— H— 2,4,6-tri-CH₃—Ph—2-CH₃—Ph— —OH (CH₃CH₂—)N— 4-Cl—Ph—(CH₃)N— H— 2,4,6-tri-CH₃—Ph—2,6-di-CH₃O—Ph— —OH H— H— PhCH₂CH₂—(CH₃)N— 4-(CH₃)₂NC(O)O— —OH H— H—CH₃(CH₂)₅—(CH₃)N— 4-(CH₃)₂NC(O)O— —OH H— H— (CH₃)₂CH—(CH₃)N—4-(CH₃)₂NC(O)O— —OH H— H— (CH₃)₃C—(CH₃)N— 4-(CH₃)₂NC(O)O— —OH H— H—(CH₃)₂CH— 4-(CH₃)₂NC(O)O— —OH (CH₃CH₂—)N— H— H— 4-pyridyl-CH₂CH₂—4-(CH₃)₂NC(O)O— —OH (CH₃)N— H— H— PhCH₂CH₂—(CH₃)N— 2,6-di-CH₃O—Ph— —OHH— H— CH₃(CH₂)₅—(CH₃)N— 2,6-di-CH₃O—Ph— —OH H— H— (CH₃)₂CH—(CH₃)N—2,6-di-CH₃O—Ph— —OH H— H— (CH₃)₃C—(CH₃)N— 2,6-di-CH₃O—Ph— —OH H— H—(CH₃)₂CH— 2,6-di-CH₃O—Ph— —OH (CH₃CH₂—)N— H— H— 4-pyridyl-CH₂CH₂—2,6-di-CH₃O—Ph— —OH (CH₃)N— cyclohexyl- H— CH₃CH₂— 4-(CH₃)₂NC(O)O— —OH(CH₃)N— H— H— CF₃CH₂— 2,6-di-CH₃O—Ph— —OH cyclohexyl- H— 2-CH₃—Ph—2,6-di-CH₃O—Ph— —OH (CH₃)N— H— H— 2-F—Ph— 2,6-di-CH₃O—Ph— —OH CH₃CH₂CH₂—H— 2-CH₃—Ph— 2,6-di-CH₃O—Ph— —OH (CH₃)N—Ph = phenyl

TABLE 5

R⁵ R⁶ R^(7′) R^(8′) R^(9′) X 4-CH₃—Ph— CH₃— H— H— 4-(CH₃)₂NC(O)O— —OH4-CH₃—Ph— CH₃— H— H— 4-(CH₃)₂NC(O)O— —OCH(CH₃)₂Ph = phenyl

TABLE 6

R⁵ R⁶ b R^(9′) X CH₃(CH₂)₅— CH₃(CH₂)₅— 2 4-HO— —OH CH₃(CH₂)₅— CH₃(CH₂)₅—2 4-(CH₃)₂NC(O)O— —OH CH₃— CH₃— 1 4-(CH₃)₂NC(O)O —OC(CH₃)₃ 3-CH₃—PhNH—H— 2 4-(CH₃)₂NC(O)O— —OH C(O)NH(CH₂)₂— CH₃(CH₂)₅— CH₃(CH₂)₅- 2 4-(1-CH₃——OH piperazin-4-yl)C(O)O—Ph = phenyl

Accordingly, the following are preferred compounds of Formulae III-IX:

-   -   N-(2-chloro-5-nitropyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-[5-(N-4-toluenesulfonylamino)pyrimidin-4-yl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester,    -   N-[5-(N-4-toluenesulfonylamino)pyrimidin-4-yl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-[5-(N-methyl-N-4-toluenesulfonylamino)pyrimidin-4-yl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester,    -   N-[5-(N-methyl-N-4-toluenesulfonylamino)pyrimidin-4-yl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-[5-(N,N-di-4-toluenesulfonylamino)pyrimidin-4-yl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-[5-[N-(1-N′-methylpyrazol-4-ylsulfonyl)-N-methylamino]pyrimidin-4-yl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-[5-(N-methyl-N-4-toluenesulfonylamino)pyrimidin-4-yl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester,    -   N-[5-(N-methyl-N-3-pyridylsulfonylamino)pyrimidin-4-yl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester,    -   N-(5-(N-methyl-N-(1-butylpyrazol-4-yl)sulfonylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(2,4-dimethoxypyrimidin-5-yl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(2,6-difluorophenyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(2-hydroxymethylphenyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N-cyclohexylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N-methyl-N-(1-methylpiperidin-4-yl)amino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N-ethyl-N-isopropylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(2,4-6-trimethylphenyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-isopropylpyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N-methyl-N-butylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbanyloxy)phenylalanine,    -   N-(2-(N-ethyl-N-propylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N,N-diethylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N-methyl-N-ethylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-benzyloxypyrimidin-4-yl)-L-phenylalanine,    -   N-(5-benzyloxypyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(N-methyl-N-4-toluenesulfonylamino)pyrimidin-4-yl)-L-phenylalanine,    -   N-(5-(N-methyl-N-3-pyridinesulfonylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-phenylpyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(3-(N-methyl-N-4-toluenesulfonylamino)pyrazin-2-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(2,2,2-trifluoroethyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(N-methyl-N-3-pyridinesulfonylamino)pyrimidin-4-yl)-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine        isopropyl ester,    -   N-(5-benzylpyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(N-methyl-N-3-pyridinesulfonylamino)pyrimidin-4-yl)-L-4-(4-methylpiperazin-N-(5-(2-trifluoromethylphenyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(2-N,N-dimethylcarbamylethyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(N-methyl-N-3-(1-methylpyrazole)sulfonylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester,    -   N-(6-phenylpyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(6-(2-trifluoromethylphenyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(6-(2-hydroxymethylphenyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-cyclohexylpyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N-methyl-N-2-furanethylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N-methyl-N-4-chlorophenylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(3-thienyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5 -(2-thienyl)pyrimidin-4-yl)-L-4-(N,N-dimethyl        carbamyloxy)phenylalanine,    -   N-(2-(N-methyl-N-2-hydroxyethylamino)-5-(2-fluorophenyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(piperidin-1-yl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(1-propylbutyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N-methyl-N-cyclobutylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N,N-bis-(2-hydroxyethyl)amino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N,N-bis-(2-hydroxyethyl)amino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N-methyl-N-phenylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(isopropoxy)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N-methyl-N-3-methylbutylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N-methylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(2-tolyl)-5        -(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N-methyl-N-2-hydroxyethylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N-methyl-N-2-methylpropylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N-methyl-N-propylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N,N-dimethylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N-methyl-N-cyclohexylamino)-5-(3-pyridyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(2-phenyl-2,2-difluoroethyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(2-phenyl-2,2-difluoroethyl)-6-chloropyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(2-phenylethyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N-methyl-N-cyclohexylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-propylpyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(2-methoxyphenyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(2-fluorophenyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N-Methyl-N-isopropylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N-isopropylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(2-phenylethyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester,    -   N-(3-(N-methyl-N-4-toluenesulfonylamino)pyrazin-2-yl)-L-phenylalanine        isopropyl ester,    -   N-(5-(2-phenylethyl)pyrimidin-4-yl)-L-phenylalanine isopropyl        ester,    -   N-(5-(N-methyl-N-3-pyridinesulfonylamino)pyrimidin-4-yl)-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine,    -   N-(2-(N-methyl-N-cyclohexylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N-methyl-N-cyclohexylamino)-5-ethylpyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        isopropyl ester,    -   N-(5-(3-nitrophenyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(3-pyridyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(2-phenylethyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-N,N-dimethylamino-5-(N-methyl-N-4-toluenesulfonylamino)pyrimidin-4-yl)-L-phenylalanine,    -   N-(5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(2-(N-methyl-N-cyclohexylamino)-5-(2-methoxyphenyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(2-(N-methyl-N-isopropylamino)-5-(2-fluorophenyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(2-(N-methyl-N-isopropylamino)-5-(2-fluorophenyl)pyrimidin-4-yl)-L-4-(2-methoxyphenyl)phenylalanine,    -   N-(2-(N-methyl-N-cyclohexylamino)-5-(2,6-difluorophenyl)pyrimidin-4-yl)-L-4-(2,6-difluorophenyl)phenylalanine,    -   N-(2-(N-methyl-N-cyclohexylamino)-5-(2-hydroxymethylphenyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(2-(N,N-bis-(2-hydroxyethyl)amino)-5-(2,4,6-trimethylphenyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(2-(N-methyl-N-cyclohexylamino)-5-(2-trifluoromethylphenyl)pyrimidin-4-yl)-L-4-(2-cyanophenyl)phenylalanine,    -   N-(2-(N-methyl-N-cyclohexylamino)-5-(3-thienyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(2-(N-methyl-N-cyclohexylamino)-5-(2-thienyl)pyrimidin-4-yl)-L-4-(4-trifluoromethylphenyl)phenylalanine,    -   N-(2-(N-methyl-N-cyclohexylamino)-5-(3-pyridyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(2-(N-methyl-N-cyclohexylamino)-5-(3-nitrophenyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(2-(N-methyl-N-cyclohexylamino)-5-(2,6-dichlorophenyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(2-(N-methyl-N-cyclohexylamino)-5-(4-pyridyl)pyrimidin-4-yl)-L-4-(3-hydroxymethylphenyl)phenylalanine,    -   N-(2-(N-ethyl-N-isopropylamino)-5-(2,6-dimethoxyphenyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(2-(N-methyl-N-cyclohexylamino)-5-(2,3-dichlorophenyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(2-(N-methyl-N-ethylamino)-5-(2,4,6-trimethylphenyl)pyrimidin-4-yl)-L-4-(2-cyanophenyl)phenylalanine,    -   N-(2-(N-methyl-N-isopropylamino)-5-(2,4,6-trimethylphenyl)pyrimidin-4-yl)-L-4-(3-pyridyl)phenylalanine,    -   N-(2-(N,N-bis-(2-hydroxyethyl)amino)-5-(2,4,6-trimethylphenyl)pyrimidin-4-yl)-L-4-(2-cyanophenyl)phenylalanine,    -   N-(2-(N-methyl-N-(1-methylpiperidin-4-yl)amino)-5-(2-cyanophenyl)pyrimidin-4-yl)-L-4-(2,6-difluorophenyl)phenylalanine,    -   N-(2-(N-ethyl-N-isopropylamino)-5-(2,4,6-trimethylphenyl)pyrimidin-4-yl)-L-4-(o-tolyl)phenylalanine,    -   N-(2-(N-methyl-N-4-chlorophenylamino)-5-(2,4,6-trimethylphenyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(5-(N-methyl-N-2-(phenyl)ethylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(N-methyl-N-hexylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(N-methyl-N-isopropylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(N-methyl-N-tert-butylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(N-ethyl-N-isopropylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(N-methyl-N-2-(4-pyridyl)ethyl-pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(5-(N-methyl-N-2-(phenyl)ethylamino)pyrimidin-4-yl)-L-4-(4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(5-(N-methyl-N-hexylamino)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(5-(N-methyl-N-isopropylamino)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(5-(N-methyl-N-tert-butylamino)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(5-(N-ethyl-N-isopropylamino)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(5-(N-methyl-N-2-(4-pyridyl)ethyl-pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(2-(N-methyl-N-cyclohexylamino)-5-ethylpyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(4-(N,N-di-n-hexylamino)-1,1-dioxo-1,2,5-thiadiazol-3-yl)-L-tyrosine,    -   N-(4-(N,N-di-n-hexylamino)-1,1-dioxo-1,2,5-thiadiazol-3-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine,    -   N-(4-(N,N-dimethylamino)-1-oxo-1,2,5-thiadiazol-3-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine        tert-butyl ester,    -   N-[4-(2-(3-methylphenylaminocarbonylamino)eth-1-ylamino)-1,1-dioxo-1,2,5-thiadiazol-3-yl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    -   N-(4-(N,N-di-n-hexylamino)-1,1-dioxo-1,2,5-thiadiazol-3-yl)-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine,    -   N-(5-(2,2,2-trifluoroethyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(2-(N-cyclohexyl-N-methyl)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(5-(2-fluorophenyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(2-(N-methyl-N-propyl)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine,    -   N-(3-chloropyrazin-2-yl)-L-4-[1-(tert-butoxycarbonyl)piperidin-4-ylcarbonylamino]phenylalanine        ethyl ester,    -   and pharmaceutically acceptable salts thereof.

Preferably, the compound is the compound of Formula O below:

Further description of the compounds of the above FormulaeIII-IX-procedures and reaction conditions for preparing these compoundsare described in U.S. Ser. No. 09/489,377 (filed Jan. 21, 2000, andissued as U.S. Pat. No. 6,492,372), herein incorporated by reference inits entirety.

Further description of these type of compounds is described in U.S.Patent Publication 20030139402, a divisional application of U.S. Ser.No. 09/489,377, herein incorporated by reference in its entirety.

Compound Preparation for Compounds of Formulae III-IX

The compounds of Formulae III-IX can be prepared from readily availablestarting materials using the following general methods and procedures.It will be appreciated that where typical or preferred processconditions (i.e., reaction temperatures, times, mole ratios ofreactants, solvents, pressures, etc.) are given, other processconditions can also be used unless otherwise stated. Optimum reactionconditions may vary with the particular reactants or solvent used, butsuch conditions can be determined by one skilled in the art by routineoptimization procedures.

Additionally, as will be apparent to those skilled in the art,conventional protecting groups may be necessary to prevent certainfunctional groups from undergoing undesired reactions. Suitableprotecting groups for various functional groups as well as suitableconditions for protecting and deprotecting particular functional groupsare well known in the art. For example, numerous protecting groups aredescribed in T. W. Greene and G. M. Wuts, Protecting Groups in OrganicSynthesis, Second Edition, Wiley, N.Y., 1991, and references citedtherein.

Furthermore, the compounds of Formulae III-IX will typically contain oneor more chiral centers. Accordingly, if desired, such compounds can beprepared or isolated as pure stereoisomers, i.e., as individualenantiomers or diastereomers, or as stereoisomer-enriched mixtures. Allsuch stereoisomers (and enriched mixtures) are included within the scopeof this invention, unless otherwise indicated. Pure stereoisomers (orenriched mixtures) may be prepared using, for example, optically activestarting materials or stereoselective reagents well-known in the art.Alternatively, racemic mixtures of such compounds can be separatedusing, for example, chiral column chromatography, chiral resolvingagents and the like.

In a preferred method of synthesis, the compounds of Formulae III-IX areprepared by coupling an amino acid derivative of the formula:

where R³ and R^(3′) are as defined herein and P¹ is a carboxylic acidprotecting group (such as an alkyl group, i.e., methyl, ethyl and thelike), with a suitably functionalized heteroaryl or heterocyclicintermediate. For example, such coupling reactions may be performed bydisplacing a leaving group, such as chloro, bromo, iodo, tosyl and thelike, from the heteroaryl or heterocyclic intermediate with the aminogroup of the amino acid derivative; or by reductive alkylation of theamino group of amino acid derivative with a carbonyl-functionalizedintermediate. Such coupling reactions are well-known to those skilled inthe art.

By way of illustration, the synthesis of a representative compound ofFormula III is shown in Scheme 1.

As shown in Scheme 1, 5-nitrouracil, 1, (commercially available fromAldrich Chemical Company, Milwaukee, Wis. USA) is treated withphosphorus oxychloride and N,N-dimethylaniline according to theprocedure described.in Whittaker, J Chem. Soc. 1951, 1565 to give1,3-dichloro-4-nitropyrimidine, 2.

1,3-Dichloro-4-nitropyrimidine, 2, is then reacted with about one molarequivalent of an amino acid derivative of the formula H₂N—CH(R³)C(O)Xwhere R³ and X are as defined herein or X is —OP¹ where P is acarboxylic acid protecting group, in the presence of a trialkylamine,such as diisopropylethylamine (DIEA). Typically, this reaction isconducted in an inert diluent, such as dichloromethane, at a temperatureranging from about 0° C. to about 10° C. for about 5 min. to about 6hours to afford intermediate 3.

The nitro group of intermediate 3is then reduced using a conventionalreducing agent, such as hydrogen and a palladium-on carbon catalyst.When hydrogen and palladium on carbon are employed as the reducingagent, the chloro group of intermediate 3 is also removed. This reactionis typically conducted by contacting 3 with a Degussa-type palladium oncarbon catalyst (typically 20%) and excess sodium bicarbonate in aninert diluent, such as methanol, under hydrogen (typically about 55 psi)for about 12 to 36 hours at ambient temperature to afford aminointermediate 4.

Amino intermediate 4 is then reacted with a sulfonyl chloride of theformula R⁵—S(O)₂—Cl, where R⁵ is as defined herein, to providesulfonamide intermediate 5. This reaction is typically conducted byreacting the amino intermediate 4 with at least one equivalent,preferably about 1.1 to about 2equivalents, of the sulfonyl chloride inan inert diluent such as dichloromethane and the like. Generally, thereaction is conducted at a temperature ranging from about −70° C. toabout 40° C. for about 1 to about 24 hours. Preferably, this reaction isconducted in the presence of a suitable base to scavenge the acidgenerated during the reaction. Suitable bases include, by way ofexample, tertiary amines, such as triethylamine, diisopropylethylamine,N-methylmorpholine and the like. Alternatively, the reaction can beconducted under Schotten-Baumann-type conditions using aqueous alkali,such as sodium hydroxide and the like, as the base. Upon completion ofthe reaction, the resulting sulfonamide 5 is recovered by conventionalmethods including neutralization, extraction, precipitation,chromatography, filtration, and the like.

Other heteroaryl intermediates may also be employed in the abovedescribed reactions including, but not limited to,2-chloro-3-nitropyrazine (J. Med. Chem. 1984, 27, 1634);4-chloro-5-nitroimidazole (J. Chem. Soc. 1930, 268); and the like.

The amino acid derivatives employed in the above reactions are eitherknown compounds or compounds that can be prepared from known compoundsby conventional synthetic procedures. For example, amino acidderivatives can be prepared by C-alkylating commercially availablediethyl 2-acetamidomalonate (Aldrich, Milwaukee, Wis., USA) with analkyl or substituted alkyl halide. This reaction is typically conductedby treating the diethyl 2-acetamidomalonate with at least one equivalentof sodium ethoxide and at least one equivalent of an alkyl orsubstituted alkyl halide in refluxing ethanol for about 6 to about 12hours. The resulting C-alkylated malonate is then deacetylated,hydrolyzed and decarboxylated by heating in aqueous hydrochloric acid atreflux for about 6 to about 12 hours to provide the amino acid,typically as the hydrochloride salt.

Examples of amino acid derivatives suitable for use in the abovereactions include, but are not limited to, L-alanine methyl ester,L-isoleucine methyl ester, L-leucine methyl ester, L-valine methylester, β-tert-butyl-L-aspartic acid methyl ester, L-asparaginetert-butyl ester, ε-Boc-L-lysine methyl ester, ε-Cbz-L-lysine methylester, γ-tert-butyl-L-glutamic acid methyl ester, L-glutamine tert-butylester, L-(N-methyl)histidine methyl-ester, L-(N-benzyl)histidine methylester, L-methionine methyl ester, L-(O-benzyl)serine methyl ester,L-tryptophan methyl ester, L-phenylalanine methyl ester, L-phenylalanineisopropyl ester, L-phenylalanine benzyl ester, L-phenylalaninamide,N-methyl-L-phenylalanine benzyl ester, 3-carboxy-D,L-phenylalaninemethyl ester, 4-carboxy-D,L-phenylalanine methyl ester,L-4-chlorophenylalanine methyl ester,L-4-(3-dimethylaminopropyloxy)-phenylalanine methyl ester,L-4-iodophenylalanine methyl ester, L-3,4-methylenedioxyphenylalaninemethyl ester, L-3,4-ethylenedioxyphenylalanine methyl ester,L-4-nitrophenylalanine methyl ester, L-tyrosine methyl ester,D,L-homophenylalanine methyl ester, L-(O-methyl)tyrosine methyl ester,L-(O-tert-butyl)tyrosine methyl ester, L-(O-benzyl)tyrosine methylester, L-3,5-diiodotyrosine methyl ester, L-3 -iodotyrosine methylester, β-(1-naphthyl)-L-alanine methyl ester, β-(2-naphthyl)-L-alaninemethyl ester, β-(2-thienyl)-L-alanine methyl ester,β-cyclohexyl-L-alanine methyl ester, β-(2-pyridyl)-L-alanine methylester, β-(3-pyridyl)-L-alanine methyl ester, β-(4-pyridyl)-L-alaninemethyl ester, β-(2-thiazolyl)-D,L-alanine methyl ester,β-(1,2,4-triazol-3-yl)-D,L-alanine methyl ester, and the like. Ifdesired, of course, other esters or amides of the above-describedcompounds may also be employed.

Additionally, α-hydroxy and α-thio carboxylic acids may also be employedin the above-described reactions. Such compounds are well-known in theart and are either commercially available or may be prepared fromcommercially available starting materials using conventional reagentsand reaction conditions.

The sulfonyl chlorides employed in the above reaction are also eitherknown compounds or compounds that can be prepared from known compoundsby conventional synthetic procedures. Such compounds are typicallyprepared from the corresponding sulfonic acid, i.e., from compounds ofthe formula R⁵—SO₃H where R⁵ is as defined above, using phosphoroustrichloride and phosphorous pentachloride. This reaction is generallyconducted by contacting the sulfonic acid with about 2to 5 molarequivalents of phosphorous trichloride and phosphorous pentachloride,either neat or in an inert solvent, such as dichloromethane, attemperature in the range of about 0° C. to about 80° C. for about 1 toabout 48 hours to afford the sulfonyl chloride. Alternatively, thesulfonyl chloride can be prepared from the corresponding thiol compound,i.e., from compounds of the formula R⁵—SH where R⁵ is as defined herein,by treating the thiol with chlorine (Cl₂) and water under conventionalreaction conditions.

Examples of sulfonyl chlorides suitable for use in this inventioninclude, but are not limited to, methanesulfonyl chloride,2-propanesulfonyl chloride, 1-butanesulfonyl chloride, benzenesulfonylchloride, 1-naphthalenesulfonyl chloride, 2-naphthalenesulfonylchloride, p-toluenesulfonyl chloride, α-toluenesulfonyl chloride,4-acetamidobenzenesulfonyl chloride, 4-amidinobenzenesulfonyl chloride,4-tert-butylbenzenesulfonyl chloride, 4-bromobenzenesulfonyl chloride,2-carboxybenzenesulfonyl chloride, 4-cyanobenzenesulfonyl chloride,3,4-dichlorobenzenesulfonyl chloride, 3,5-dichlorobenzenesulfonylchloride, 3,4-dimethoxybenzenesulfonyl chloride,3,5-ditrifluoromethylbenzenesulfonyl chloride, 4-fluorobenzenesulfonylchloride, 4-methoxybenzenesulfonyl chloride,2-methoxycarbonylbenzenesulfonyl chloride, 4-methylamidobenzenesulfonylchloride, 4-nitrobenzenesulfonyl chloride, 4-thioamidobenzenesulfonylchloride, 4-trifluoromethylbenzenesulfonyl chloride,4-trifluoromethoxybenzenesulfonyl chloride,2,4,6-trimethylbenzenesulfonyl chloride, 2-phenylethanesulfonylchloride, 2-thiophenesulfonyl chloride, 5-chloro-2-thiophenesulfonylchloride, 2,5-dichloro-4-thiophenesulfonyl chloride, 2-thiazolesulfonylchloride, 2-methyl-4-thiazolesulfonyl chloride,1-methyl-4-imidazolesulfonyl chloride, 1-methyl-4-pyrazolesulfonylchloride, 5-chloro-1,3-dimethyl-4-pyrazolesulfonyl chloride,3-pyridinesulfonyl chloride, 2-pyrimidinesulfonyl chloride and the like.If desired, a sulfonyl fluoride, sulfonyl bromide or sulfonic acidanhydride may be used in place of the sulfonyl chloride in the abovereaction to form the sulfonamide intermediate 5.

If desired, sulfonamide intermediate 5 can be alkylated at thesulfonamide nitrogen atom to provide compound 6. For example, 5 can becontacted with excess diazomethane (generated, for example, using1-methyl-3-nitro-1-nitrosoguanidine and sodium hydroxide) to afford 6where R⁶ is methyl. Other conventional alkylation procedures andreagents may also be employed to prepare various compounds of thisinvention.

In another preferred embodiment, compounds of Formulae III-IX may beprepared by displacement of a leaving group as shown in Scheme 2:

where R³, Q and X are as defined herein; A′ is heteroaryl, substitutedheteroaryl, heterocyclic or substituted heterocyclic containing twonitrogen atoms in the heteroaryl or heterocyclic ring; and L¹ is aleaving group, such as chloro, bromo, iodo, sulfonate ester and thelike.

Typically, this reaction is conducted by combining approximatelystoichiometric equivalents of 7 and 8 in a suitable inert diluent suchas water, dimethylsulfoxide (DMSO) and the like, with an excess of asuitable base such as sodium bicarbonate, sodium hydroxide, etc. toscavenge the acid generated by the reaction. The reaction is preferablyconducted at from about 25° C. to about 100° C. until reactioncompletion which typically occurs within 1 to about 24 hours. Thisreaction is further described in U.S. Pat. No. 3,598,859, which isincorporated herein by reference in its entirety. Upon reactioncompletion, the product 9 is recovered by conventional methods includingprecipitation, chromatography, filtration and the like.

In still another alternative embodiment, compounds of Formulae III-IX inwhich Q is NR⁴ can be prepared by reductive amination of a suitable2-oxocarboxylic acid ester, 10, such as a pyruvate ester, as shown inScheme 3:

where A′, R³ and X are as defined herein.

Generally, this reaction is conducted by combining equamolar amounts of10 and 11 in an inert diluent such as methanol, ethanol and the likeunder conditions which provide for imine formation (not shown). Theimine formed is then reduced under conventional conditions by a suitablereducing agent such as sodium cyanoborohydride, H₂/palladium on carbonand the like to form the product 12. In a particularly preferredembodiment, the reducing agent is H₂/palladium on carbon which isincorporated into the initial reaction medium thereby permitting iminereduction in situ in a one pot procedure to provide 12. The reaction ispreferably conducted at from about 20° C. to about 80° C. at a pressureof from 1to atmospheres until reaction completion which typically occurswithin 1to about 24 hours. Upon reaction completion, the product 12 isrecovered by conventional methods including chromatography, filtrationand the like.

Alternatively, certain compounds of Formulae III-IX can be prepared viaa rhodium-catalyzed insertion reaction as shown in Scheme 4:

where A″ is heteroaryl or substituted heteroaryl containing two nitrogenatoms in the heteroaryl ring, and R³ and X (preferably alkoxy) are asdefined herein. Typically, this reaction is conducted using rhodiumacetate dimer, Rh₂(OAc)₄, in an inert diluent such as toluene at atemperature ranging from about 25° C. to about 80° C. for about 1 to 12hours to afford 15. This reaction is described further in B. R. Henkeet. al., J. Med. Chem. 1998, 41, 5020-5036 and references cited therein.

Similarly, certain compounds of Formulae III-IX can be prepared by thecopper-catalyzed coupling reaction shown in Scheme 5:

where A″ is as defined herein, X³ is halogen, such as chloro, bromo oriodo (preferably iodo), and R³ and X (preferably alkoxy) are as definedherein. Typically, this reaction is conducted using copper iodide (CuI)and potassium carbonate in an inert diluent such as N,N-dimethylacetamide (DMA) at a temperature ranging from about 60° C. to about 120°C. for about 12 to 36 hours to afford 15. This reaction is describedfurther in D. Ma et. al., J. Am. Chem. Soc. 1998, 120, 12459-12467 andreferences cited therein.

For ease of synthesis, the compounds of Formulae III-IX are typicallyprepared as an ester, i.e., where X is an alkoxy or substituted alkoxygroup and the like. If desired, the ester group can be hydrolysed usingconventional conditions and reagents to provide the correspondingcarboxylic acid. Typically, this reaction is conducted by treating theester with at least one equivalent of an alkali metal hydroxide, such aslithium, sodium or potassium hydroxide, in an inert diluent, such asmethanol or mixtures of methanol and water, at a temperature rangingabout 0° C. to about 24° C. for about 1 to about 12 hours.Alternatively, benzyl esters may be removed by hydrogenolysis using apalladium catalyst, such as palladium on carbon, and tert-butyl esterscan be removed using formic acid to afford the corresponding carboxylicacid.

As will be apparent to those skilled in the art, other functional groupspresent on any of the substituents of the compounds of Formulae III-IXcan be readily modified or derivatized either before or after theabove-described synthetic reactions using well-known syntheticprocedures. For example, a nitro group present on a substituent of acompound of Formulae III-IX or an intermediate thereof may be readilyreduced by hydrogenation in the presence of a palladium catalyst, suchas palladium on carbon, to provide the corresponding amino group. Thisreaction is typically conducted at a temperature of from about 20° C. toabout 50° C. for about 6 to about 24 hours in an inert diluent, such asmethanol. Compounds having a nitro group on the R³ and/or R^(3′)substituent can be prepared, for example, by using a4-nitrophenylalanine derivative and the like in the above-describedcoupling reactions.

Similarly, a pyridyl group can be hydrogenated in the presence of aplatinum catalyst, such as platinum oxide, in an acidic diluent toprovide the corresponding piperidinyl analogue. Generally, this reactionis conducted by treating the pyridine compound with hydrogen at apressure ranging from about 20 psi to about 60 psi, preferably about 40psi, in the presence of the catalyst at a temperature of about 20° C. toabout 50° C. for about 2to about 24 hours in an acidic diluent, such asa mixture of methanol and aqueous hydrochloric acid.

Additionally, when the R³ and/or R^(3′) substituent of a compound ofFormulae III-IX or an intermediate thereof contains a primary orsecondary amino group, such amino groups can be further derivatizedeither before or after the above coupling reactions to provide, by wayof example, amides, sulfonamides, ureas, thioureas, carbamates,secondary or tertiary amines and the like. Compounds having a primaryamino group on the R³ and/or R^(3′) substituent may be prepared, forexample, by reduction of the corresponding nitro compound as describedabove.

By way of illustration, a compound of Formulae III-IX or an intermediatethereof having a substituent containing a primary or secondary aminogroup, such as where R³ is a (4-aminophenyl)methyl group, can be readilyN-acylated using conventional acylating reagents and conditions toprovide the corresponding amide. This acylation reaction is typicallyconducted by treating the amino compound with at least one equivalent,preferably about 1.1 to about 1.2 equivalents, of a carboxylic acid inthe presence of a coupling reagent such as a carbodiimide, BOP reagent(benzotriazol-1-yloxy-tris(dimethylamino)phosphoniumhexafluorophosphonate) and the like, in an inert diluent, such asdichloromethane, chloroform, acetonitrile, tetrahydrofuran,N,N-dimethylformamide and the like, at a temperature ranging from about0° C. to about 37° C. for about 4 to about 24 hours. Preferably, apromoter, such as N-hydroxysuccinimide, 1-hydroxy-benzotriazole and thelike, is used to facilitate the acylation reaction. Examples ofcarboxylic acids suitable for use in this reaction include, but are notlimited to, N-tert-butyloxycarbonylglycine,N-tert-butyloxycarbonyl-L-phenylalanine,N-tert-butyloxycarbonyl-L-aspartic acid benzyl ester, benzoic acid,N-tert-butyloxycarbonylisonipecotic acid, N-methylisonipecotic acid,N-tert-butyloxycarbonylnipecotic acid,N-tert-butyloxycarbonyl-L-tetrahydroisoquinoline-3-carboxylic acid,N-(toluene-4-sulfonyl)-L-proline and the like.

Alternatively, a compound of Formulae III-IX or an intermediate thereofcontaining a primary or secondary amino group can be N-acylated using anacyl halide or a carboxylic acid anhydride to form the correspondingamide. This reaction is typically conducted by contacting the aminocompound with at least one equivalent, preferably about 1.1 to about 1.2equivalents, of the acyl halide or carboxylic acid anhydride in an inertdiluent, such as dichloromethane, at a temperature ranging from about−70° C. to about 40° C. for about 1to about 24 hours. If desired, anacylation catalyst such as 4-(N,N-dimethylamino)pyridine may be used topromote the acylation reaction. The acylation reaction is preferablyconducted in the presence of a suitable base to scavenge the acidgenerated during the reaction. Suitable bases include, by way ofexample, tertiary amines, such as triethylamine, diisopropylethylamine,N-methylmorpholine and the like. Alternatively, the reaction can beconducted under Schotten-Baumann-type conditions using aqueous alkali,such as sodium hydroxide and the like.

Examples of acyl halides and carboxylic acid anhydrides suitable for usein this reaction include, but are not limited to, 2-methylpropionylchloride, trimethylacetyl chloride, phenylacetyl chloride, benzoylchloride, 2-bromobenzoyl chloride, 2-methylbenzoyl chloride,2-trifluoro-methylbenzoyl chloride, isonicotinoyl chloride, nicotinoylchloride, picolinoyl chloride, acetic anhydride, succinic anhydride, andthe like. Carbamyl chlorides, such as N,N-dimethylcarbamyl chloride,N,N-diethylcarbamyl chloride and the like, can also be used in thisreaction to provide ureas. Similarly, dicarbonates, such asdi-tert-butyl dicarbonate, may be employed to provide carbamates.

In a similar manner, a compound of Formulae III-IX or an intermediatethereof containing a primary or secondary amino group may beN-sulfonated to form a sulfonamide using a sulfonyl halide or a sulfonicacid anhydride. Sulfonyl halides and sulfonic acid anhydrides suitablefor use in this reaction include, but are not limited to,methanesulfonyl chloride, chloromethanesulfonyl chloride,p-toluenesulfonyl chloride, trifluoromethanesulfonic anhydride, and thelike. Similarly, sulfamoyl chlorides, such as dimethylsulfamoylchloride, can be used to provide sulfamides (e.g., >N—SO₂—N<).

Additionally, a primary and secondary amino group present on asubstituent of a compound of Formulae III-IX or an intermediate thereofcan be reacted with an isocyanate or a thioisocyanate to give a urea orthiourea, respectively. This reaction is typically conducted bycontacting the amino compound with at least one equivalent, preferablyabout 1.1 to about 1.2 equivalents, of the isocyanate or thioisocyanatein an inert diluent, such as toluene and the like, at a temperatureranging from about 24° C. to about 37° C. for about 12 to about 24hours. The isocyanates and thioisocyanates used in this reaction arecommercially available or can be prepared from commercially availablecompounds using well-known synthetic procedures. For example,isocyanates and thioisocyanates are readily prepared by reacting theappropriate amine with phosgene or thiophosgene. Examples of isocyanatesand thioisocyanates suitable for use in this reaction include, but arenot limited to, ethyl isocyanate, n-propyl isocyanate, 4-cyanophenylisocyanate, 3-methoxyphenyl isocyanate, 2-phenylethyl isocyanate, methylthioisocyanate, ethyl thioisocyanate, 2-phenylethyl thioisocyanate,3-phenylpropyl thioisocyanate, 3-(N,N-diethylamino)propylthioisocyanate, phenyl thioisocyanate, benzyl thioisocyanate, 3-pyridylthioisocyanate, fluorescein isothiocyanate (isomer I) and the like.

Furthermore, when a compound of Formulae-III-IX or an intermediatethereof contains a primary or secondary amino group, the amino group canbe reductively alkylated using aldehydes or ketones to form a secondaryor tertiary amino group. This reaction is typically conducted bycontacting the amino compound with at least one equivalent, preferablyabout 1.1 to about 1.5 equivalents, of an aldehyde or ketone and atleast one equivalent based on the amino compound of a metal hydridereducing agent, such as sodium cyanoborohydride, in an inert diluent,such as methanol, tetrahydrofuran, mixtures thereof and the like, at atemperature ranging from about 0° C. to about 50° C. for about 1to about72 hours. Aldehydes and ketones suitable for use in this reactioninclude, by way of example, benzaldehyde, 4-chlorobenzaldehyde,valeraldehyde and the like.

In a similar manner, when a compound of Formulae III-IX or anintermediate thereof has a substituent containing a hydroxyl group, thehydroxyl group can be further modified or derivatized either before orafter the above coupling reactions to provide, by way of example,ethers, carbamates and the like. Compounds having a hydroxyl group onthe R³ substituent, for example, can be prepared using an amino acidderivative derived from tyrosine and the like in the above-describedreactions.

By way of example, a compound of Formulae III-IX or an intermediatethereof having a substituent containing a hydroxyl group, such as whereR³ is a (4-hydroxyphenyl)methyl group, can be readily O-alkylated toform ethers. This O-alkylation reaction is typically conducted bycontacting the hydroxy compound with a suitable alkali or alkaline earthmetal base, such as potassium carbonate, in an inert diluent, such asacetone, 2-butanone and the like, to form the alkali or alkaline earthmetal salt of the hydroxyl group. This salt is generally not isolated,but is reacted in situ with at least one equivalent of an alkyl orsubstituted alkyl halide or sulfonate, such as an alkyl chloride,bromide, iodide, mesylate or tosylate, to afford the ether. Generally,this reaction is conducted at a temperature ranging from about 60° C. toabout 150° C. for about 24 to about 72 hours. Preferably, a catalyticamount of sodium or potassium iodide is added to the reaction mixturewhen an alkyl chloride or bromide is employed in the reaction.

Examples of alkyl or substituted alkyl halides and sulfonates suitablefor use in this reaction include, but are not limited to, tert-butylbromoacetate, N-tert-butyl chloroacetamide, 1-bromoethylbenzene, ethylα-bromophenylacetate, 2-(N-ethyl-N-phenylamino)ethyl chloride,2-(N-N-ethylamino)ethyl chloride, 2-(N,N-diisopropylamino)ethylchloride, 2-(N,N-dibenzylamino)ethyl chloride, 3-(N,N-ethylamino)propylchloride, 3-(N-benzyl-N-methylamino)propyl chloride,N-(2-chloroethyl)morpholine, 2-(hexamethyleneimino)ethyl chloride,3-(N-methylpiperazine)propyl chloride,1-(3-chlorophenyl)-4-(3-chloropropyl)piperazine,2-(4-hydroxy-4-phenylpiperidine)ethyl chloride,N-tert-butyloxycarbonyl-3-piperidinemethyl tosylate, and the like.

Alternatively, a hydroxyl group present on a substituent of a compoundof Formulae III-IX or an intermediate thereof can be O-alkylating usingthe Mitsunobu reaction. In this reaction, an alcohol, such as3-(N,N-dimethylamino)-1-propanol and the like, is reacted with about 1.0to about 1.3 equivalents of triphenylphosphine and about 1.0 to about1.3 equivalents of diethyl azodicarboxylate in an inert diluent, such astetrahydrofuran, at a temperature ranging from about −10° C. to about 5°C. for about 0.25 to about 1hour. About 1.0 to about 1.3 equivalents ofa hydroxy compound, such as N-tert-butyltyrosine methyl ester, is thenadded and the reaction mixture is stirred at a temperature of about 0°C. to about 30° C. for about 2 to about 48 hours to provide theO-alkylated product.

In a similar manner, a compound of Formulae III-IX or an intermediatethereof containing an aryl hydroxy group can be reacted with an aryliodide to provide a diaryl ether. Generally, this reaction is conductedby forming the alkali metal salt of the hydroxyl group using a suitablebase, such as sodium hydride, in an inert diluent such as xylenes at atemperature of about −25° C. to about 10° C. The salt is then treatedwith about 1.1 to about 1.5 equivalents of cuprous bromide dimethylsulfide complex at a temperature ranging from about 10° C. to about 30°C. for about 0.5 to about 2.0 hours, followed by about 1.1 to about 1.5equivalents of an aryl iodide, such as sodium 2-iodobenzoate and thelike. The reaction is then heated to about 70° C. to about 150° C. forabout 2 to about 24 hours to provide the diaryl ether.

Additionally, a hydroxy-containing compound can also be readilyderivatized to form a carbamate. In one method for preparing suchcarbamates, a hydroxy compound of Formulae III-IX or an intermediatethereof is contacted with about 1.0 to about 1.2 equivalents of4-nitrophenyl chloroformate in an inert diluent, such asdichloromethane, at a temperature ranging from about −25° C. to about 0°C. for about 0.5 to about 2.0 hours. Treatment of the resultingcarbonate with an excess, preferably about 2 to about 5 equivalents, ofa trialkylamine, such as triethylamine, for about 0.5 to 2 hours,followed by about 1.0 to about 1.5 equivalents of a primary or secondaryamine provides the carbamate. Examples of amines suitable for using inthis reaction include, but are not limited to, piperazine,1-methylpiperazine, 1-acetylpiperazine, morpholine, thiomorpholine,pyrrolidine, piperidine and the like.

Alternatively, in another method for preparing carbamates, ahydroxy-containing compound is contacted with about 1.0 to about 1.5equivalents of a carbamyl chloride in an inert diluent, such asdichloromethane, at a temperature ranging from about 25° C. to about 70°C. for about 2 to about 72 hours. Typically, this reaction is conductedin the presence of a suitable base to scavenge the acid generated duringthe reaction. Suitable bases include, by way of example, tertiaryamines, such as triethylamine, diisopropylethylamine, N-methylmorpholineand the like. Additionally, at least one equivalent (based on thehydroxy compound) of 4-(N,N-dimethylamino)pyridine is preferably addedto the reaction mixture to facilitate the reaction. Examples of carbamylchlorides suitable for use in this reaction include, by way of example,dimethylcarbamyl chloride, diethylcarbamyl chloride and the like.

Likewise, when a compound of Formulae III-IX or an intermediate thereofcontains a primary or secondary hydroxyl group, such hydroxyl groups canbe readily converted into a leaving group and displaced to form, forexample, amines, sulfides and fluorides. Generally, when a chiralcompound is employed in these reactions, the stereochemistry at thecarbon atom attached to the derivatized hydroxyl group is typicallyinverted.

These reactions are typically conducted by first converting the hydroxylgroup into a leaving group, such as a tosylate, by treatment of thehydroxy compound with at least one equivalent of a sulfonyl halide, suchas p-toluenesulfonyl chloride and the like, in pyridine. This reactionis generally conducted at a temperature of from about 0° C. to about 70°C. for about 1to about 48 hours. The resulting tosylate can then bereadily displaced with sodium azide, for example, by contacting thetosylate with at least one equivalent of sodium azide in an inertdiluent, such as a mixture of N,N-dimethylformamide and water, at atemperature ranging from about 0° C. to about 37° C. for about 1to about12 hours to provide the corresponding azido compound. The azido groupcan then be reduced by, for example, hydrogenation using a palladium oncarbon catalyst to provide the amino (—NH₂) compound.

Similarly, a tosylate group can be readily displaced by a thiol to forma sulfide. This reaction is typically conducted by contacting thetosylate with at least one equivalent of a thiol, such as thiophenol, inthe presence of a suitable base, such as 1,8diazabicyclo[5.4.0]undec-7-ene (DBU), in an inert diluent, such asN,N-dimethylformamide, at a temperature of from about 0° C. to about 37°C. for about 1to about 12 hours to provide the sulfide. Additionally,treatment of a tosylate with morpholinosulfur trifluoride in an inertdiluent, such as dichloromethane, at a temperature ranging from about 0°C. to about 37° C. for about 12 to about 24 hours affords thecorresponding fluoro compound.

Furthermore, a compound of Formulae III-IX or an intermediate thereofhaving a substituent containing an iodoaryl group, for example, when R³is a (4-iodophenyl)methyl group, can be readily converted either beforeor after the above coupling reactions into a biaryl compound. Typically,this reaction is conducted by treating the iodoaryl compound with about1.1 to about 2 equivalents of an arylzinc iodide, such as2-(methoxycarbonyl)phenylzinc iodide, in the presence of a palladiumcatalyst, such as palladium tetra(triphenylphosphine), in an inertdiluent, such as tetrahydrofuran, at a temperature ranging from about24° C. to about 30° C. until reaction completion. This reaction isfurther described, for example, in Rieke, J. Org. Chem. 1991, 56, 1445.Additional methods for preparing biaryl derivatives are disclosed inInternational Publication Number WO 98/53817, published Dec. 3, 1998,the disclosure of which is incorporated herein by reference in itsentirety.

In some cases, the compounds of Formulae III-IX or intermediates thereofmay contain substituents having one or more sulfur atoms. When present,such sulfur atoms can be oxidized either before or after the abovecoupling reactions to provide a sulfoxide or sulfone compound usingconventional reagents and reaction conditions. Suitable reagents foroxidizing a sulfide compound to a sulfoxide include, by way of example,hydrogen peroxide, 3-chloroperoxybenzoic acid (MCPBA), sodium periodateand the like. The oxidation reaction is typically conducted bycontacting the sulfide compound with about 0.95 to about 1.1 equivalentsof the oxidizing reagent in an inert diluent, such as dichloromethane,at a temperature ranging from about −50° C. to about 75° C. for about1to about 24 hours. .The resulting sulfoxide can then be furtheroxidized to the corresponding sulfone by contacting the sulfoxide withat least one additional equivalent of an oxidizing reagent, such ashydrogen peroxide, MCPBA, potassium permanganate and the like.Alternatively, the sulfone can be prepared directly by contacting thesulfide with at least two equivalents, and preferably an excess, of theoxidizing reagent. Such reactions are described further in March,“Advanced Organic Chemistry,” 4th Ed., pp. 1201-1202, Wiley Publisher,1992.

Other procedures and reaction conditions for preparing the compounds ofthis invention are described in the examples set forth below.Additionally, other procedures for preparing compounds useful in certainaspects of this invention are disclosed in U.S. Ser. No. 09/489,378,filed on Jan. 21, 2000, entitled “Compounds Which Inhibit LeucocyteAdhesion Mediated by VLA-4,” now issued as U.S. Pat. No. 6,479,492, thedisclosure of which is incorporated herein by reference in its entirety.

Compounds of Formulae X-XV

In one aspect, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compounds ofFormulae X, XI, XII, XIII, XIV, and XV.

In one aspect, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compoundsdefined by Formula X below. These compounds have a binding affinity toVLA-4 as expressed by an IC₅₀ of about 15 μM or less (measured asdescribed in Example A below):

-   -   wherein each X is independently fluoro, chloro or bromo;    -   p is an integer from 0 to 3;    -   R¹ and R³ together with the nitrogen atom to which they are        bound form an azetidinyl, pyrrolidinyl, pyrrolyl,        2,5-dihydopyrrol-1-yl, piperidinyl, or        1,2,3,6-tetrahydropyridin-1-yl;    -   R² is selected from the group consisting of lower alkyl, lower        aikenyi, and lower alkylenecycloalkyl;    -   and pharmaceutically acceptable salts thereof.

In a preferred embodiment, R¹ and R³ together with the nitrogen atom towhich they are bound form an azetidinyl, pyrrolidinyl, or piperidinylgroup.

In one aspect, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compoundsdefined by Formula XI below. These compounds have a binding affinity toVLA-4 as expressed by an IC₅₀ of about 15 μM or less (measured asdescribed in Example A below):

-   -   wherein each X is independently selected from the group        consisting of fluoro and chloro;    -   m is an integer equal to 1or 2;    -   R² is selected from the group consisting of lower alkyl, lower        alkenyl, and lower alkylenecycloalkyl;    -   R¹ and R³ together with the nitrogen atom to which they are        bound form an azetidinyl, pyrrolidinyl, or piperidinyl group;    -   and pharmaceutically acceptable salts thereof.

In one aspect, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compoundsdefined by Formula XII below. These compounds have a binding affinity toVLA-4 as expressed by an IC₅₀ of about 15 μM or less (measured asdescribed in Example A below):

-   -   wherein each X is independently fluoro or chloro;    -   n is zero or one;    -   R² is —CH₂—R′ where R′ is selected from the group consisting of        hydrogen, methyl or —CH═CH₂;    -   R¹ and R³ together with the nitrogen atom to which they are        bound form an azetidinyl, pyrrolidinyl, or piperidinyl group;    -   and pharmaceutically acceptable salts thereof.

In one aspect, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compoundsdefined by Formula XIII below. These compounds have a binding affinityto VLA-4 as expressed by an IC₅₀ of about 15 μM or less (measured asdescribed in Example A below):

-   -   wherein each X is independently fluoro, chloro or bromo;    -   p is an integer from 0 to 3;    -   R¹ and R³ together with the nitrogen atom to which they are        bound form an azetidinyl, pyrrolidinyl, pyrrolyl,        2,5-dihydopyrrol-1-yl, piperidinyl, or        1,2,3,6-tetrahydropyridin-1-yl;    -   R² is lower alkynyl;    -   and pharmaceutically acceptable salts thereof.

In a preferred embodiment, R¹ and R³ together with the nitrogen atom towhich they are bound form an azetidinyl, pyrrolidinyl, or piperidinylgroup and R² is propargyl.

In one aspect, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compoundsdefined by Formula XIV below. These compounds have a binding affinity toVLA-4 as expressed by an ICso of about 15 μM or less (measured asdescribed in Example A below):

-   -   wherein each X is independently selected from the group        consisting of fluoro and chloro;    -   m is an integer equal to 1 or 2;    -   R² is lower alkynyl;    -   R¹ and R³ together with the nitrogen atom to which they are        bound form an azetidinyl, pyrrolidinyl, or piperidinyl group;    -   and pharmaceutically acceptable salts thereof.

In one aspect, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compoundsdefined by Formula XV below. These compounds have a binding affinity toVLA-4 as expressed by an IC₅₀ of about 15 μM or less (measured asdescribed in Example A below):

-   -   wherein each X is independently fluoro or chloro;    -   n is zero or one;    -   R² is lower alkynyl;    -   R¹ and R³ together with the nitrogen atom to which they are        bound form an azetidinyl, pyrrolidinyl, or piperidinyl group;    -   and pharmaceutically acceptable salts thereof.

N-[2-N′,N′-diethylamino-5-aminosulfonylphenylpyrimidin-4-yl]-p-carbomyloxy-phenylalaninecompounds within the scope of this invention include those set forth inTable 7 as follows: TABLE 7

R¹ and R³ R²

Example No. pyrrolidinyl ethyl 4-chlorophenyl 505 pyrrolidinyl ethyl4-fluorophenyl 506 pyrrolidinyl methyl 4-fluorophenyl 507 pyrrolidinylmethyl 4-chlorophenyl 508 piperidinyl methyl 4-fluorophenyl 509piperidinyl ethyl 4-fluorophenyl 510 azetidinyl ethyl 4-fluorophenyl 511azetidinyl methyl 4-fluorophenyl 512 azetidinyl methyl 4-chlorophenyl513 azetidinyl ethyl 4-chlorophenyl 514 pyrrolidinyl methyl2,4-difluorophenyl 515 pyrrolidinyl ethyl 2,4-difluorophenyl 516azetidinyl methyl 2,4-difluorophenyl 517 azetidinyl ethyl2,4-difluorophenyl 518 pyrrolidinyl propargyl 4-fluorophenyl 519pyrrolidinyl progargyl 2,4-difluorophenyl 520 azetidinyl propargyl2,4-difluorophenyl 521 azetidinyl propargyl 4-fluorophenyl 522pyrrolidinyl progargyl 4-chlorophenyl 523

Specific compounds within the scope of this invention include thefollowing compounds. As used below, these compounds are named based onphenylalanine derivatives but, alternatively, these compounds could havebeen named based onN-[2-N′,N′-diethylamino-5-aminosulfonylphenyl-pyrimidin-4-yl]-p-carbomyloxyphenylalaninederivatives or2-{2-diethylamino-5-[(benzenesulfonyl)methylamino]-pyrimidin-4-ylamino}-p-carbamoyloxy-phenyl)propionicacid derivatives.

-   -   N-(2-[N′,N′-diethylamino]-5-[N″-(4-chlorophenylsulfonyl)-N″-ethylamino]pyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalanine;    -   N-(2-[N′,N′-diethylamino]-5-[N″-(4-fluorophenylsulfonyl)-N″-ethylamino]pyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalanine;    -   N-(2-[N′,N′-diethylamino]-5-[N″-(4-fluorophenylsulfonyl)-N″-methylamino]pyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalanine;    -   N-(2-[N′,N′-diethylamino]-5-[N″-(4-chlorophenylsulfonyl)-N″-methylamino]pyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalanine;    -   N-(2-[N′,N′-diethylamino]-5-[N″-(4-fluorophenylsulfonyl)-N″-methylamino]pyrimidin-4-yl)-4′-(piperidin-1-ylcarbonyloxy)-L-phenylalanine;    -   N-(2-[N′,N′-diethylamino]-5-[N″-(4-fluorophenylsulfonyl)-N″-ethylamino]pyrimidin-4-yl)-4′-(piperidin-1-ylcarbonyloxy)-L-phenyl        alanine;    -   N-(2-[N′,N′-diethylamino]-5-[N″-(4-fluorophenylsulfonyl)-N″-ethylamino]pyrimidin-4-yl)-4′-(azetidin-1-ylcarbonyloxy)-L-phenylalanine;    -   N-(2-[N′,N′-diethylamino]-5-[N″-(4-fluorophenylsulfonyl)-N″-methylamino]pyrimidin-4-yl)-4′-(azetidin-1-ylcarbonyloxy)-L-phenylalanine;    -   N-(2-[N′,N′-diethylamino]-5-[N″-(4-chlorophenylsulfonyl)-N″-methylamino]pyrimidin-4-yl)-4′-(azetidin-1-ylcarbonyloxy)-L-phenylalanine;    -   N-(2-[N′,N′-diethylamino]-5-[N″-(4-chlorophenylsulfonyl)-N″-ethylamino]pyrimidin-4-yl)-4′-(azetidin-1-ylcarbonyloxy)-L-phenylalanine;    -   N-(2-[N′,N′-diethylamino]-5-[N″-(2,4-difluorophenylsulfonyl)-N″-methylamino]pyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalanine;    -   N-(2-[N′,N′-diethylamino]-5-[N″-(2,4-difluorophenylsulfonyl)-N″-ethylamino]pyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalanine;    -   N-(2-[N′,N′-diethylamino]-5-[N″-(2,4-difluorophenylsulfonyl)-N″-methylamino]pyrimidin-4-yl)-4′-(azetidin-1-ylcarbonyloxy)-L-phenylalanine;    -   N-(2-[N′,N′-diethylamino]-5-[N″-(2,4-difluorophenylsulfonyl)-N″-ethylamino]pyrimidin-4-yl)-4′-(azetidin-1-ylcarbonyloxy)-L-phenylalanine;    -   N-(2-[N′,N′-diethylamino]-5-[N″-(4-fluorophenylsulfonyl)-N″-propargylamino]pyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalanine;    -   N-(2-[N′,N′-diethylamino]-5-[N″-(2,4-difluorophenylsulfonyl)-N″-propargylamino]pyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalanine;    -   N-(2-[N′,N′-diethylamino]-5-[N″-(2,4-difluorophenylsulfonyl)-N″-propargylamino]pyrimidin-4-yl)-4′-(azetidin-1-ylcarbonyloxy)-L-phenylalanine;    -   N-(2-[N′,N′-diethylamino]-5-[N″-(4-fluorophenylsulfonyl)-N″-propargylamino]pyrimidin-4-yl)-4′-(azetidin-1-ylcarbonyloxy)-L-phenylalanine;    -   N-(2-[N′,N′-diethylamino]-5-[N″-(4-chlorophenylsulfonyl)-N″-propargylamino]pyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalanine;        and    -   pharmaceutically acceptable salts thereof.

Preferably, the compound is the compound of Formula P below:

In another embodiment, preferably the compound is the compound ofFormula Q below:

Compound Preparation of Compounds of Formulae X-XV

The compounds of Formulae X-XV can be prepared from readily availablestarting materials using the methods and procedures set forth in theexamples below. These methods and procedures outline specific reactionprotocols for preparingN-[2-N′,N′-diethylamino-5-aminosulfonylphenyl-yrimidin-4-yl]-p-carbomyloxy-phenylalaninecompounds. Compounds within the scope not exemplified in these examplesand methods are readily prepared by appropriate substitution of startingmaterials which are either commercially available or well known in theart.

Other procedures and reaction conditions for preparing the compounds ofthis invention are described in the examples set forth below.Additionally, other procedures for preparing compounds useful in certainaspects of this invention are disclosed in U.S. Pat. 6,492,372, issuedDec. 10, 2002; the disclosure of which is incorporated herein byreference in its entirety.

Compounds of Formulae XVI-XXI

In one aspect, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compounds ofFormulae XVI, XVII, XVIII, XIX, XX, and XXI.

In one aspect, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compoundsdefined by Formula XVI below. These compounds have a binding affinity toVLA-4 as expressed by an IC₅₀ of about 15 μM or less (measured asdescribed in Example A below):

-   -   wherein each X is independently fluoro, chloro or bromo;    -   p is 0 or an integer from 1-3;    -   R¹ is selected from the group consisting of methyl and ethyl;    -   R² is selected from the group consisting of lower alkyl, lower        alkenyl, and lower alkylenecycloalkyl;    -   and pharmaceutically acceptable salts thereof.

In one aspect, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compoundsdefined by Formula XVII below. These compounds have a binding affinityto VLA-4 as expressed by an ICso of about 15 μM or less (measured asdescribed in Example A below):

-   -   wherein each X is independently selected from the group        consisting of fluoro and chloro,    -   m is an integer equal to 1 or 2;    -   R² is selected from the group consisting of lower alkyl, lower        alkenyl, and lower alkylenecycloalkyl;    -   and pharmaceutically acceptable salts thereof.

In one aspect, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compoundsdefined by Formula XVIII below. These compounds have a binding affinityto VLA-4 as expressed by an IC₅₀ of about 15 μM or less (measured asdescribed in Example A below):

-   -   wherein each X is independently fluoro or chloro;    -   n is zero or one;    -   R² is —CH₂—R′ where R′ is selected from the group consisting of        hydrogen, methyl or —CH═CH₂;    -   and pharmaceutically acceptable salts thereof.

In one aspect, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compoundsdefined by Formula XIX below. These compounds have a binding affinity toVLA-4 as expressed by an IC₅₀ of about 15 μM or less (measured asdescribed in Example A below):

-   -   wherein each X is independently fluoro, chloro or bromo;    -   p is 0 or an integer from 1-3;    -   R¹ is selected from the group consisting of methyl and ethyl;    -   R² is lower alkynyl;    -   and pharmaceutically acceptable salts thereof.

In one aspect, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compoundsdefined by Formula XX below. These compounds have a binding affinity toVLA-4 as expressed by an IC₅₀ of about 15 μM or less (measured asdescribed in Example A below):

-   -   wherein each X is independently selected from the group        consisting of fluoro and chloro,    -   m is an integer equal to 1 or 2;    -   R² is lower alkynyl;    -   and pharmaceutically acceptable salts thereof.

In one aspect, the compounds that can be utilized as combinationtherapies with methotrexate for the treatment of RA are compoundsdefined by Formula XXI below. These compounds have a binding affinity toVLA-4 as expressed by an IC₅₀ of about 15 μM or less (measured asdescribed in Example A below):

-   -   wherein each X is independently fluoro or chloro;    -   n is zero or one;    -   R² is lower alkynyl;    -   and pharmaceutically acceptable salts thereof.    -   R² is preferably propargyl in any of one of Formula XIX, XX or        XXI.

N-[2-N′,N′-diethylamino-5-aminosulfonylphenylpyrimidin-4-yl]-p-carbomyloxyphenylalaninecompounds within the scope of this invention include those set forth inTable 8 as follows: TABLE 8 XVII

Example No.

R² 524 4-fluorophenyl methyl 525 4-chlorophenyl methyl 5263,4-difluorophenyl methyl 527 3,4-dichlorophenyl methyl 528 phenylmethyl 529 2-fluorophenyl methyl 530 3-fluorophenyl methyl 5314-fluorophenyl isopropyl 532 4-fluorophenyl ethyl 533 3,4-difluorophenylisopropyl 534 4-chlorophenyl isopropyl 535 3,4-difluorophenyl ethyl 5364-chlorophenyl ethyl 537 4-fluorophenyl cyclopropylmethyl 5383,5-difluorophenyl methyl 539 3,5-difluorophenyl ethyl 5402,4-difluorophenyl methyl 541 2,4-difluorophenyl ethyl 5423,5-dichlorophenyl methyl 543 3,5-dichlorophenyl ethyl 5444-fluorophenyl n-propyl 545 4-fluorophenyl allyl 546 4-fluorophenylisobutyl 547 4-fluorophenyl n-butyl 548 2,6-difluorophenyl Methyl 5492,3-difluorophenyl methyl 550 4-fluorophenyl propargyl 5512,4-difluorophenyl propargyl 552 4-fluorophenyl 2-trisfluoroethyl

Specific compounds within the scope of this invention include thefollowing. As used below, these compounds are named based on propionicacid derivatives but, alternatively, these compounds could have beennamed based onN-[2-N′,N′-diethylamino-5-aminosulfonylphenylpyrimidin-4-yl]-p-carbomyloxy-phenylalaninederivatives.

-   -   1-{2-diethylamino-5-[(4-chlorobenzenesulfonyl)methylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5[(4-fluorobenzenesulfonyl)methylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(3,4-difluorobenzenesulfonyl)methylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(3,4-dichlorobenzenesulfonyl)methylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(benzenesulfonyl)methylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(2-fluorobenzenesulfonyl)methylamino]-pyrimisin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(3-fluorobenzenesulfonyl)methylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(4-fluorobenzenesulfonyl)isopropylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(4-fluorobenzenesulfonyl)ethylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(3,4-difluorobenzenesulfonyl)isopropylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(4-chlorobenzenesulfonyl)isopropylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(3,4-difluorobenzenesulfonyl)ethylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(4-chlorobenzenesulfonyl)ethylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(4-fluorobenzenesulfonyl)cylclopropylmethyl-amino]pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(3,5-difluorobenzenesulfonyl)methylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(3,5-difluorobenzenesulfonyl)ethylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(2,4-difluorobenzenesulfonyl)methylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(2,4-difluorobenzenesulfonyl)ethylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(3,5-dichlorobenzenesulfonyl)methylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(3,5-dichlorobenzenesulfonyl)ethylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(4-fluorobenzenesulfonyl)-n-propylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(4-fluorobenzenesulfonyl)allylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(4-fluorobenzenesulfonyl)isobotylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(4-fluorobenzenesulfonyl)-n-butylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(2,6-difluorobenzenesulfonyl)methylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-diethylamino-5-[(2,3-difluorobenzenesulfonyl)ethylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-Diethylamino-5-[(4-fluorobenzenesulfonyl)propargylamino]pyrimidin-4-ylamino        }-3-(4-dimethylcarbamoyloxyphenyl)propionic acid;    -   2-{2-Diethylamino-5-[(2,4-difluorobenzenesulfonyl)propargylamino]pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   2-{2-Diethylamino-5-[(4-fluorobenzenesulfonyl)-(2-trisfluoroethyl)-amino]pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic        acid;    -   and pharmaceutically acceptable salts thereof        Compound Preparation for Compounds of Formulae XVI-XXI

The compounds of Formulae XVI-XXI can be prepared from readily availablestarting materials using the methods and procedures set forth in theexamples below. These methods and procedures outline specific reactionprotocols for preparingN-[2-N′,N+-diethylamino-5-aminosulfonylphenyl-yrimidin-4-yl]-p-carbomyloxy-phenylalaninecompounds. Compounds within the scope not exemplified in these examplesand methods are readily prepared by appropriate substitution of startingmaterials which are either commercially available or well known in theart.

Other procedures and reaction conditions for preparing the compounds ofthis invention are described in the examples set forth below.Additionally, other procedures for preparing compounds useful in certainaspects of this invention are disclosed in U.S. Pat. No. 6,492,372 thedisclosure of which is incorporated herein by reference in its entirety.

Pharmaceutical Formulations of the Compounds

In general, the compounds of the subject invention will be administeredin a therapeutically effective amount by any of the accepted modes ofadministration for these compounds. The compounds can be administered bya variety of routes, including, but not limited to, oral, parenteral(e.g., subcutaneous, subdural, intravenous, intramuscular, intrathecal,intraperitoneal, intracerebral, intraarterial, or intralesional routesof administration), topical, intranasal, localized (e.g., surgicalapplication or surgical suppository), rectal, and pulmonary (e.g.,aerosols, inhalation, or powder). Accordingly, these compounds areeffective as both injectable and oral compositions. The compounds can beadministered continuously by infusion or by bolus injection. Preferably,the compounds are administered by parenteral routes. More preferably,the compounds are administered by intravenous routes. Such compositionsare prepared in a manner well known in the pharmaceutical art.

The actual amount of the compound of the subject invention, i.e., theactive ingredient, will depend on a number of factors, such as theseverity of the disease, i.e., the condition or disease to be treated,age and relative health of the subject, the potency of the compoundused, the route and form of administration, and other factors.

Toxicity and therapeutic efficacy of such compounds can be determined bystandard pharmaceutical procedures in cell cultures or experimentalanimals, e.g., for determining the LD₅₀ (the dose lethal to 50% of thepopulation) and the ED₅₀ (the dose therapeutically effective in 50% ofthe population). The dose ratio between toxic and therapeutic effects isthe therapeutic index and it can be expressed as the ratio LD₅₀/ED₅₀.Compounds that exhibit large therapeutic indices are preferred.

The data obtained from the cell culture assays and animal studies can beused in formulating a range of dosage for use in humans. The dosage ofsuch compounds lies preferably within a range of circulatingconcentrations that include the ED₅₀ with little or no toxicity. Thedosage may vary within this range depending upon the dosage formemployed and the route of administration utilized. For any compound usedin the method of the invention, the therapeutically effective dose canbe estimated initially from cell culture assays. A dose may beformulated in animal models to achieve a circulating plasmaconcentration range which includes the IC₅₀ (i.e., the concentration ofthe test compound which achieves a half-maximal inhibition of symptoms)as determined in cell culture. Such information can be used to moreaccurately determine useful doses in humans. Levels in plasma may bemeasured, for example, by high performance liquid chromatography. Theeffective blood level of the compounds of the subject invention ispreferably greater than or equal to 10 ng/ml.

The amount of the pharmaceutical composition administered to the patientwill vary depending upon what is being administered, the purpose of theadministration, such as prophylaxis or therapy, the state of thepatient, the manner of administration, and the like. In therapeuticapplications, compositions are administered to a patient alreadysuffering from a disease in an amount sufficient to cure or at leastpartially arrest the symptoms of the disease and its complications. Anamount adequate to accomplish this is defined as “therapeuticallyeffective dose.” Amounts effective for this use will depend on thedisease condition being treated as well as by the judgment of theattending clinician depending upon factors such as the severity of theinflammation, the age, weight and general condition of the patient, andthe like.

The compositions administered to a patient are in the form ofpharmaceutical compositions described supra. These compositions may besterilized by conventional sterilization techniques, or may be sterilefiltered. The resulting aqueous solutions may be packaged for use as is,or lyophilized, the lyophilized preparation being combined with asterile aqueous carrier prior to administration. The pH of the compoundpreparations fypically will be between 3 and 11, more preferably from 5to 9 and most preferably from 7 to 8. It will be understood that use ofcertain of the foregoing excipients, carriers, or stabilizers willresult in the formation of pharmaceutical salts.

The active compound is effective over a wide dosage range and isgenerally administered in a pharmaceutically or therapeuticallyeffective amount. The therapeutic dosage of the compounds of the presentinvention will vary according to, for example, the particular use forwhich the treatment is made, the manner of administration of thecompound, the health and condition of the patient, and the judgment ofthe prescribing physician. For example, for intravenous administration,the dose will typically be in the range of about 0.5 mg to about 100 mgper kilogram body weight, preferably about 3 mg to about 50 mg perkilogram body weight. Effective doses can be extrapolated fromdose-response curves derived from in vitro or animal model test systems.Typically, the clinician will administer the compound until a dosage isreached that achieves the desired effect.

According to one aspect of the invention, the compounds are administeredin combination with methotrexate, to treat, ameliorate, or palliate thesymptoms of rheumatoid arthritis. When administered in combination, thecompounds may be administered in the same formulation as themethotrexate, or in a separate formulation. The compounds may beadministered prior to, following, or concurrently with the methotrexatesuch that the benefitsd of the combination therapy are achieved. Thecalculation of appropriate dosages will be well within the purvue of theskilled artisan. Standard doses of methotrexate for the treatment ofrheumatoid arthritis range from 2 mg to 20 mg per dose per week. Dosagesof the compounds are as set forth above. The methotrexate dosage may beadministered as a single dose or as a divided dose. Once a response hasbeen achieved, the dosage may be reduced if possible to the lowesteffective dose. The maximum recommended dose is 20 mg/week. Preferably,methotrexate is administered orally or via injection.

When employed as pharmaceuticals, the compounds of the subject inventionare usually administered in the form of pharmaceutical compositions.This invention also includes pharmaceutical compositions, which containas the active ingredient, one or more of the compounds of the subjectinvention above, associated with one or more pharmaceutically acceptablecarriers or excipients. The excipient employed is typically one suitablefor administration to human subjects or other mammals. In making thecompositions of this invention, the active ingredient is usually mixedwith an excipient, diluted by an excipient or enclosed within a carrierwhich can be in the form of a capsule, sachet, paper or other container.When the excipient serves as a diluent, it can be a solid, semi-solid,or liquid material, which acts as a vehicle, carrier or medium for theactive ingredient. Thus, the compositions can be in the form of tablets,pills, powders, lozenges, sachets, cachets, elixirs, suspensions,emulsions, solutions, syrups, aerosols (as a solid or in a liquidmedium), ointments containing, for example, up to 10% by weight of theactive compound, soft and hard gelatin capsules, suppositories, sterileinjectable solutions, and sterile packaged powders.

In preparing a formulation, it may be necessary to mill the activecompound to provide the appropriate particle size prior to combiningwith the other ingredients. If the active compound is substantiallyinsoluble, it ordinarily is milled to a particle size of less than 200mesh. If the active compound is substantially water soluble, theparticle size is normally adjusted by milling to provide a substantiallyuniform distribution in the formulation, e.g., about 40 mesh.

Some examples of suitable excipients include lactose, dextrose, sucrose,sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates,tragacanth, gelatin, calcium silicate, microcrystalline cellulose,polyvinylpyrrolidone, cellulose, sterile water, syrup, and methylcellulose. The formulations can additionally include: lubricating agentssuch as talc, magnesium stearate, and mineral oil; wetting agents;emulsifying and suspending agents; preserving agents such as methyl- andpropylhydroxy-benzoates; sweetening agents; and flavoring agents. Thecompositions of the invention can be formulated so as to provide quick,sustained or delayed release of the active ingredient afteradministration to the patient by employing procedures known in the art.

The quantity of active compound in the pharmaceutical composition andunit dosage form thereof may be varied or adjusted widely depending uponthe particular application, the manner or introduction, the potency ofthe particular compound, and the desired concentration. The term “unitdosage forms” refers to physically discrete units suitable as unitarydosages for human subjects and other mammals, each unit containing apredetermined quantity of active material calculated to produce thedesired therapeutic effect, in association -with asuitable-pharmaceutical excipient. The concentration of therapeuticallyactive compound may vary from about 1 mg/ml to 250 g/ml.

Preferably, the compound can be formulated for parenteral administrationin a suitable inert carrier, such as a sterile physiological salinesolution. For example, the concentration of compound in the carriersolution is typically between about 1-100 mg/ml. The dose administeredwill be determined by route of administration. Preferred routes ofadministration include parenteral or intravenous administration. Atherapeutically effective dose is a dose effective to produce asignificant steroid tapering. Preferably, the amount is sufficient toproduce a statistically significant amount of steroid tapering in asubject.

Administration of therapeutic agents by intravenous formulation is wellknown in the pharmaceutical industry. An intravenous formulation shouldpossess certain qualities aside from being just a composition in whichthe therapeutic agent is soluble. For example, the formulation shouldpromote the overall stability of the active ingredient(s), also, themanufacture of the formulation should be cost effective. All of thesefactors ultimately determine the overall success and usefulness of anintravenous formulation.

Other accessory additives that may be included in pharmaceuticalformulations of compounds of the present invention as follow: solvents:ethanol, glycerol, propylene glycol; stabilizers: EDTA (ethylene diaminetetraacetic acid), citric acid; antimicrobial preservatives: benzylalcohol, methyl paraben, propyl paraben; buffering agents: citricacid/sodium citrate, potassium hydrogen tartrate, sodium hydrogentartrate, acetic acid/sodium acetate, maleic acid/sodium maleate, sodiumhydrogen phthalate, phosphoric acid/potassium dihydrogen phosphate,phosphoric acid/disodium hydrogen phosphate; and tonicity modifiers:sodium chloride, mannitol, dextrose.

The presence of a buffer is necessary to maintain the aqueous pH in therange of from about 4 to about 8 and more preferably in a range of fromabout 4 to about 6. The buffer system is generally a mixture of a weakacid and a soluble salt thereof, e.g., sodium citrate/citric acid; orthe monocation or dication salt of a dibasic acid, e.g., potassiumhydrogen tartrate; sodium hydrogen tartrate, phosphoric acid/potassiumdihydrogen phosphate, and phosphoric acid/disodium hydrogen phosphate.

The amount of buffer system used is dependent on (1) the desired pH; and(2) the amount of drug. Generally, the amount of buffer used is in a0.5:1 to 50:1 mole ratio of buffer:alendronate (where the moles ofbuffer are taken as the combined moles of the buffer ingredients, e.g,sodium citrate and citric acid) of formulation to maintain a pH in therange of 4 to 8 and generally, a 1:1 to 10:1 mole ratio of buffer(combined) to drug present is used.

A useful buffer in the invention is sodium citrate/citric acid in therange of 5 to 50 mg per ml. sodium citrate to 1 to 15 mg per ml. citricacid, sufficient to maintain an aqueous pH of 4-6 of the composition.

The buffer agent may also be present to prevent the precipitation of thedrug through soluble metal complex formation with dissolved metal ions,e.g., Ca, Mg, Fe, Al, Ba, which may leach out of glass containers orrubber stoppers or be present in ordinary tap water. The agent may actas a competitive complexing agent with the drug and produce a solublemetal complex leading to the presence of undesirable particulates.

In addition, the presence of an agent, e.g., sodium chloride in anamount of about of 1-8 mg/ml, to adjust the tonicity to the same valueof human blood may be required to avoid the swelling or shrinkage oferythrocytes upon administration of the intravenous formulation leadingto undesirable side effects such as nausea or diarrhea and possibly toassociated blood disorders. In general, the tonicity of the formulationmatches that of human blood which is in the range of 282 to 288 mOsm/kg,and in general is 285 mOsm/kg, which is equivalent to the osmoticpressure corresponding to a 0.9% solution of sodium chloride.

The intravenous formulation can be administered by direct intravenousinjection, i.v. bolus, or can be administered by infusion by addition toan appropriate infusion solution such as 0.9% sodium chloride injectionor other compatible infusion solution.

The compositions are preferably formulated in a unit dosage form, eachdosage containing from about 5 to about 100 mg, more usually about 10 toabout 30 mg, of the active ingredient. The term “unit dosage forms”refers to physically discrete units suitable as unitary dosages forhuman subjects and other mammals, each unit containing a predeterminedquantity of active material calculated to produce the desiredtherapeutic effect, in association with a suitable pharmaceuticalexcipient.

The active compound is effective over a wide dosage range and isgenerally administered in a pharmaceutically effective amount. It, willbe understood, however, that the amount of the compound actuallyadministered will be determined by a physician, in the light of therelevant circumstances, including the condition to be treated, thechosen route of administration, the actual compound administered, theage, weight, and response of the individual patient, the severity of thepatient's symptoms, and the like.

For preparing solid compositions such as tablets, the principal activeingredient is mixed with a pharmaceutical excipient to form a solidpreformulation composition containing a homogeneous mixture of acompound of the present invention. When referring to thesepreformulation compositions as homogeneous, it is meant that the activeingredient is dispersed evenly throughout the composition so that thecomposition may be readily subdivided into equally effective unit dosageforms such as tablets, pills and capsules. This solid preformulation isthen subdivided into unit dosage forms of the type described abovecontaining from, for example, 0.1 to about 500 mg of the activeingredient of the present invention.

The tablets or pills of the present invention may be coated or otherwisecompounded to provide a dosage form affording the advantage of prolongedaction. For example, the tablet or pill can comprise an inner dosage andan outer dosage component, the latter being in the form of an envelopeover the former. The two components can be separated by an enteric layerwhich serves to resist disintegration in the stomach and permit theinner component to pass intact into the duodenum or to be delayed inrelease. A variety of materials can be used for such enteric layers orcoatings, such materials including a number of polymeric acids andmixtures of polymeric acids with such materials as shellac, cetylalcohol, and cellulose acetate.

The liquid forms in which the novel compositions of the presentinvention may be incorporated for administration orally or by injectioninclude aqueous solutions suitably flavored syrups, aqueous or oilsuspensions, and flavored emulsions with edible oils such as cottonseedoil, sesame oil, coconut oil, or peanut oil, as well as elixirs andsimilar pharmaceutical vehicles.

Compositions for inhalation or insufflation include solutions andsuspensions in pharmaceutically acceptable, aqueous or organic solvents,or mixtures thereof, and powders. The liquid or solid compositions maycontain suitable pharmaceutically acceptable excipients as describedsupra. Preferably the compositions are administered by the oral or nasalrespiratory route for local or systemic effect. Compositions inpreferably pharmaceutically acceptable solvents may be nebulized by useof inert gases. Nebulized solutions may be breathed directly from thenebulizing device or the nebulizing device may be attached to a facemasks tent, or intermittent positive pressure breathing machine.Solution, suspension, or powder compositions may be administered,preferably orally or nasally, from devices which deliver the formulationin an appropriate manner.

The compounds of this invention can be administered in a sustainedrelease form. Suitable examples of sustained-release preparationsinclude semipermeable matrices of solid hydrophobic polymers containingthe protein, which matrices are in the form of shaped articles, e.g.,films, or microcapsules. Examples of sustained-release matrices includepolyesters, hydrogels (e.g., poly(2-hydroxyethyl-methacrylate) asdescribed by Langer et al., J. Biomed. Mater. Res. 15: 167-277 (1981)and Langer, Chem. Tech. 12: 98-105 (1982) or poly(vinyl alcohol)),polylactides (U.S. Pat. No. 3,773,919), copolymers of L-glutamic acidand gamma ethyl-L-glutamate (Sidman et al., Biopolymers 22: 547-556,1983), non-degradable ethylene-vinyl acetate (Langer et al., supra),degradable lactic acid-glycolic acid copolymers such as the LUPRONDEPOT™ (i.e., injectable microspheres composed of lactic acid-glycolicacid copolymer and leuprolide acetate), and poly-D-(−)-3-hydroxybutyricacid (EP 133,988).

The compounds of this invention can be administered in a sustainedrelease form, for example a depot injection, implant preparation, orosmotic pump, which can be formulated in such a manner as to permit asustained release of the active ingredient. Implants for sustainedrelease formulations are well-known in the art. Implants may beformulated as, including but not limited to, microspheres, slabs, withbiodegradable or non-biodegradable polymers. For example, polymers oflactic acid and/or glycolic acid form an erodible polymer that iswell-tolerated by the host. The implant is placed in proximity to thesite of protein deposits (e.g., the site of formation of amyloiddeposits associated with neurodegenerative disorders), so that the localconcentration of active agent is increased at that site relative to therest of the body.

The following formulation examples illustrate pharmaceuticalcompositions of the present invention.

FORMULATION EXAMPLE 1

Hard gelatin capsules containing the following ingredients are prepared:Quantity Ingredient (mg/capsule) Active Ingredient 30.0 Starch 305.0Magnesium stearate 5.0

The above ingredients are mixed and filled into hard gelatin capsules in340 mg quantities.

FORMULATION EXAMPLE 2

A tablet formula is prepared using the ingredients below: QuantityIngredient (mg/capsule) Active Ingredient 25.0 Cellulose,microcrystalline 200.0 Colloidal silicon dioxide 10.0 Stearic acid 5.0

The components are blended and compressed to form tablets, each weighing240 mg.

FORMULATION EXAMPLE 3

A dry powder inhaler formulation is prepared containing the followingcomponents: Ingredient Weight % Active Ingredient 5 Lactose 95

The active mixture is mixed with the lactose and the mixture is added toa dry powder inhaling appliance.

FORMULATION EXAMPLE 4

Tablets, each containing 30 mg of active ingredient, are prepared asfollows: Quantity Ingredient (mg/capsule) Active Ingredient 30.0 mgStarch 45.0 mg Microcrystalline cellulose 35.0 mg Polyvinylpyrrolidone(as 10% solution in water) 4.0 mg Sodium carboxymethyl starch 4.5 mgMagnesium stearate 0.5 mg Talc 1.0 mg Total 120 mg

The active ingredient, starch and cellulose are passed through a No. 20mesh U.S. sieve and mixed thoroughly. The solution ofpolyvinyl-pyrrolidone is mixed with the 15 resultant powders, which arethen passed through a 16 mesh U.S. sieve. The granules so produced aredried at 50° to 60° C. and passed through a 16 mesh U.S. sieve. Thesodium carboxymethyl starch, magnesium stearate, and talc, previouslypassed through a No.30 mesh U.S. sieve, are then added to the granules,which after mixing, are compressed on a tablet machine to yield tabletseach weighing 150 mg.

FORMULATION EXAMPLE 5

Capsules, each containing 40 mg of medicament are made as follows:Quantity Ingredient (mg/capsule) Active Ingredient 40.0 mg Starch 109.0mg Magnesium stearate 1.0 mg Total 150.0 mg

The active ingredient, cellulose, starch, an magnesium stearate areblended, passed through a No. 20 mesh U.S. sieve, and filled into hardgelatin capsules in 150 mg quantities.

FORMULATION EXAMPLE 6

Suppositories, each containing 25 mg of active ingredient are made asfollows: Ingredient Amount Active Ingredient 25 mg Saturated fatty acidglycerides to 2,000 mg

The active ingredient is passed through a No. 60 mesh U.S. sieve andsuspended in the saturated fatty acid glycerides previously melted usingthe minimum heat necessary. The mixture is then poured into asuppository mold of nominal 2.0 g capacity and allowed to cool.

FORMUATION EXAMPLE 7

Suspensions, each containing 50 mg of medicament per 5.0 ml dose aremade as follows: Ingredient Amount Active Ingredient 50.0 mg Xanthan gum4.0 mg Sodium carboxymethyl cellulose (11%) 50.0 mg Microcrystallinecellulose (89%) Sucrose 1.75 g Sodium benzoate 10.0 mg Flavor and Colorq.v. Purified water to 5.0 ml

The medicament, sucrose and xanthan gum are blended, passed through aNo. 10 mesh U.S. sieve, and then mixed with a previously made solutionof the microcrystalline cellulose and sodium carboxymethyl cellulose inwater. The sodium benzoate, flavor, and color are diluted with some ofthe water and added with stirring. Sufficient water is then added toproduce the required volume.

FORMULATION EXAMPLE 8

Hard gelatin tablets, each containing 15 mg of active ingredient aremade as follows: Quantity Ingredient (mg/capsule) Active Ingredient 15.0mg Starch 407.0 mg Magnesium stearate 3.0 mg Total 425.0 mg

The active ingredient, cellulose, starch, and magnesium stearate areblended, passed through a No. 20 mesh U.S. sieve, and filled into hardgelatin capsules in 560 mg quantities.

FORMULATION EXAMPLE 9

An intravenous formulation may be prepared as follows: IngredientQuantity Active Ingredient 250.0 mg Isotonic saline 1000 ml

Therapeutic compound compositions generally are placed into a containerhaving a sterile access port, for example, an intravenous solution bagor vial having a stopper pierceable by a hypodermic injection needle orsimilar sharp instrument.

FORMULATION EXAMPLE 10

A topical formulation may be prepared as follows: Ingredient QuantityActive Ingredient 1-10 g Emulsifying Wax 30 g Liquid Paraffin 20 g WhiteSoft Paraffin to 100 g

The white soft paraffin is heated until molten. The liquid paraffin andemulsifying wax are incorporated and stirred until dissolved. The activeingredient is added and stirring is continued until dispersed. Themixture is then cooled until solid.

FORMULATION EXAMPLE 11

An aerosol formulation may be prepared as follows:

A solution of the candidate compound in 0.5% sodium bicarbonate/saline(w/v) at a concentration of 30.0 mg/mL is prepared using the followingprocedure:

A. Preparation of 0.5% Sodium Bicarbonate/Saline Stock Solution: 100.0mL Ingredient Gram/100.0 mL Final Concentration Sodium Bicarbonate 0.5 g0.5% Saline q.s. ad 100.0 mL q.s. ad 100%

Procedure:

-   -   1. Add 0.5g sodium bicarbonate into a 100 mL volumetric flask.    -   2. Add approximately 90.0 mL saline and sonicate until        dissolved.    -   3. Q.S. to 100.0 mL with saline and mix-thoroughly.

B. Preparation of 30.0 mg/mL Candidate Compound: 10.0 mL IngredientGram/10.0 mL Final Concentration Candidate Compound 0.300 g 30.0 mg/mL0.5% Sodium Bicarbonate/ q.s. ad 10.0 mL q.s ad 100% Saline StockSolution

Procedure:

-   -   1. Add 0.300 g of the candidate compound into a 10.0 mL        volumetric flask.    -   2. Add approximately 9.7 mL of 0.5% sodium bicarbonate/saline        stock solution.    -   3. Sonicate until the candidate compound is completely        dissolved.    -   4. Q.S. to 10.0 mL with 0.5% sodium bicarbonate/saline stock        solution and mix thoroughly.

Another preferred formulation employed in the methods of the presentinvention employs transdermal delivery devices (“patches”). Suchtransdermal patches may be used to provide continuous or discontinuousinfusion of the compounds of the present invention in controlledamounts. The construction and use of transdermal patches for thedelivery of pharmaceutical agents is well known in the art. See, e.g.,U.S. Pat. No. 5,023,252, issued Jun. 11, 1991, herein incorporated byreference. Such patches may be constructed for continuous, pulsatile, oron demand delivery of pharmaceutical agents.

Direct or indirect placement techniques may be used when it is desirableor necessary to introduce the pharmaceutical composition to the brain.Direct techniques usually involve placement of a drug delivery catheterinto the host′s ventricular system to bypass the blood-brain barrier.One such implantable delivery system used for the transport ofbiological factors to specific anatomical regions of the body isdescribed in U.S. Pat. No. 5,011,472, which is herein incorporated byreference.

Indirect techniques, which are generally preferred, usually involveformulating the compositions to provide for drug latentiation by theconversion of hydrophilic drugs into lipid-soluble drugs. Latentiationis generally achieved through blocking of the hydroxy, carbonyl,sulfate, and primary amine groups present on the drug to render the drugmore lipid soluble and amenable to transportation across the blood-brainbarrier. Alternatively, the delivery of hydrophilic drugs may beenhanced by intra-arterial infusion of hypertonic solutions which cantransiently open the blood-brain barrier.

According to one aspect of the invention, the compound may beadministered alone with methotrexate, as a combination of compounds andwith methotrexate, or in combination with anti-alpha-4-antibodies andmethotrexate. The compounds of the present invention may also beadministered in combination with an immunosuppressant, wherein theimmunosuppressant is typically used to treat the condition or diseasefor which the compound of the present invention is being administered.The immunosuppressant may b,e, but is not limited to, azathioprine,6-mercaptopurine, or mycophenolate. When administered in combination,the small compounds may be administered in the same formulation as theseother compounds or compositions, or in a separate formulation. Whenadministered in combinations, the compounds may be administered priorto, following, or concurrently with the other compounds andmethotrexate.

Pharmaceutical compositions of the invention are suitable for use in avariety of drug delivery systems. Suitable formulations for use in thepresent invention are found in Remington's Pharmaceutical Sciences, MacePublishing Company, Philadelphia, Pa., 17th ed. (1985).

In order to enhance serum half-life, the compounds may be encapsulated,introduced into the lumen of liposomes, prepared as a colloid, or otherconventional techniques may be employed which provide an extended serumhalf-life of the compounds. A variety of methods are available forpreparing liposomes, as described in, e.g., Szoka et al., U.S. Pat. Nos.4,235,871, 4,501,728 and 4,837,028 each of which is incorporated hereinby reference.

Polymer Conjugates

Compounds of the present invention may be formulated and administered aspolymer conjugates, preferably PEG derivatives. Polymer conjugates mayexhibit benefits over non-conjugated polymers, such as improvedsolubility and stability.

As such, single polymer molecules may be employed for conjugation withthe compounds of the present invention, although it is also contemplatedthat more than one polymer molecule can be attached as well. Theconjugated compounds of the present invention may find utility in bothin vivo as well as non-in vivo applications. Additionally, it will berecognized that the conjugating polymer may utilize any other groups,moieties, or other conjugated species, as appropriate to the end useapplication. By way of example, it may be useful in some applications tocovalently bond to the polymer a functional moiety impartingUV-degradation resistance, or antioxidation, or other properties orcharacteristics to the polymer. As a further example, it may beadvantageous in some applications to functionalize the polymer to renderit reactive and enable it to cross-link to a drug molecule and toenhance various properties or characteristics of the overall conjugatedmaterial. Accordingly, the polymer may contain any functionality,repeating groups, linkages, or other constitutent structures which donot preclude the efficacy of the conjugated the compounds of the presentinvention composition for its intended purpose.

Illustrative polymers that may usefully be employed to achieve thesedesirable characteristics are described supra, as well as in PCT WO01/54690 (to Zheng et al.) incorporated by reference herein in itsentirety. The polymer may be coupled to the compounds of the presentinvention (preferably via a linker moiety) to form stable bonds that arenot significantly cleavable by human enzymes. Generally, for a bond tobe not “significantly” cleavable requires that no more than about 20% ofthe bonds connecting the polymer and the compounds of the presentinvention to which the polymer is linked, are cleaved within a 24 hourperiod, as measured by standard techniques in the art including, but notlimited to, high pressure liquid chromatography (HPLC).

The compounds of the present inventions are conjugated most preferablyvia a terminal reactive group on the polymer although conjugations canalso be branched from non-terminal reactive groups. The polymer with thereactive group(s) is designated herein as “activated polymer”. Thereactive group selectively reacts with reactive groups on the compoundsof the present invention. The activated polymer(s) is reacted so thatattachment may occur at any available functional group on compounds ofthe present invention. Amino, carbon, free carboxylic groups, suitablyactivated carbonyl groups, hydroxyl, guanidyl, oxidized carbohydratemoieties, amino, carbon and mercapto groups of the compounds of thepresent invention (if available) can be used as attachment sites.

Generally, about 1.0 to about 10 moles of activated polymer per mole ofthe compounds of the present invention, depending on concentration, isemployed. The final amount is a balance between maximizing the extent ofthe reaction while minimizing non-specific modifications of the productand, at the same time, defining chemistries that will maintain optimumactivity, while at the same time optimizing the half-life of thecompounds of the present invention. Preferably, at least about 50% ofthe biological activity of the compounds of the present invention isretained, and most preferably 100% is retained.

The reactions may take place by any suitable art-recognized method usedfor reacting biologically active materials with inert polymers.Generally, the process involves preparing an activated polymer andthereafter reacting the compounds of the present invention with theactivated polymer to produce a soluble compound suitable forformulation. This modification reaction can be performed by severalmethods, which may involve one or more steps. The polymeric substancesincluded herein are preferably water-soluble at room temperature. Anon-limiting list of such polymers includes polyalkylene oxidehomopolymers such as polyethylene glycol (PEG) or polypropylene glycols,polyoxyethylenated polyols, copolymers thereof and block copolymersthereof, provided that the water solubility of the block copolymers ismaintained.

In the preferred practice of the present invention, polyalkylene glycolresidues of C₁-C₄ alkyl polyalkylene glycols, preferably polyethyleneglycol (PEG), or poly(oxy)alkylene glycol residues of such glycols areadvantageously incorporated in the polymer systems of interest. Thus,the polymer to which the compounds of the present invention are attachedmay be a homopolymer of polyethylene glycol (PEG) or is apolyoxyethylated polyol, provided in all cases that the polymer issoluble in water at room temperature. Non-limiting examples of suchpolymers include polyalkylene oxide homopolymers such as PEG orpolypropylene glycols, polyoxyethylenated glycols, copolymers thereofand block copolymers thereof, provided that the water solubility of theblock copolymer is maintained.

Examples of polyoxyethylated polyols include, but are not limited to,polyoxyethylated glycerol, polyoxyethylated sorbitol, polyoxyethylatedglucose, or the like. The glycerol backbone of polyoxyethylated glycerolis the same backbone occurring naturally in, for example, animals andhumans in mono-, di-, and triglycerides. Therefore, this branching wouldnot necessarily be seen as a foreign agent in the body.

Those of ordinary skill in the art will recognize that the foregoinglist is merely illustrative and that all polymer materials having thequalities described herein are contemplated. The polymer need not haveany particular molecular weight, but it is preferred that the molecularweight be between about 300 and 100,000, more preferably between 10,000and 40,000. In particular, sizes of 20,000 or more are most effective atpreventing loss of the product due to filtration in the kidneys.

Polyethylene glycol (PEG) and related polyalkylene oxides (PAOs) areknown in the art as being useful adjuncts for the preparation of drugs.See for example, PCT WO 93/24476. PEG has also been conjugated toproteins, peptides and enzymes to increase aqueous solubility andcirculating life in vivo as well as reduce antigenicity. See, forexample, U.S. Pat. Nos. 5,298,643 and 5,321,095, both to Greenwald etal. PCT WO 93/24476 discloses using an ester linkage to covalently bindan organic molecule to water-soluble polyethylene glycols. Thus, thecompounds of the invention are preferably administered as polyethyleneglycol (PEG) derivatives. Further description of polyethylene glycolderivatives of the compounds of the present invention and reactionconditions for preparing these derivatives are described in U.S. Ser.No. 60/538,573, entitled “Polyethylene Glycol Conjugates of Dipeptides,”filed Jan. 23, 2004, herein incorporated by reference in its entirety.

As such, the compounds or conjugates of this invention may contain oneor more polyethylene glycol (PEG) substituents covalently attachedthereto. Such conjugates demonstrate improved serum half-life, ascompared to compounds lacking polyethylene glycol substituents. Withoutbeing limited to any theory, the improved serum half-life is believed tobe associated with the covalent conjugation of at least one polyethyleneglycol entity onto the structure of the compound.

The term “PEG” refers to polymers comprising multiple oxyalkylene units.Such polymers are optionally mono-capped with a substituent preferablyselected from alkyl, aryl, substituted alkyl, and substituted aryl.Inclusive of such polymers are those diamino capped polyoxyalkylenepolymers which are known in the art as Jeffamines®. Still further, suchpolymers can optionally contain one or more non-oxyalkylene units suchas the commercially available poly[di(ethylene glycol)adipates,poly[di(ethylene glycol)phthalate diols, and the like.

By PEG derivative is meant a polyethylene glycol polymer in which one orboth of the terminal hydroxyl groups found in polyethylene glycol itselfhas been modified. Examples of suitable modifications include replacingone or both hydroxyl group(s) with alternative functional groups, whichmay be protected or unprotected, with low molecular weight ligands, orwith another macromolecule or polymer. Modification of the terminalhydroxyl groups in the polyethylene glycol may be achieved by reactingthe polyethylene glycol with compounds comprising complementary reactivefunctional groups, including functional groups which are able to undergoa reaction with the hydroxyl groups in polyethylene glycol. The PEGderivatives of the compounds of this invention may contain one or morepolyethylene glycol (PEG) substituents covalently attached thereto by alinking group.

“Linking group” or “linker” refers to a group or groups that covalentlylinks a non-PEG substituted compound of the present invention with oneor more PEG groups. Each linker may be chiral or achiral, linear,branched or cyclic and may be homogenous or heterogeneous in its atomcontent (e.g., linkers containing only carbon atoms or linkerscontaining carbon atoms as well as one or more heteroatoms present onthe linker.

The PEG group or groups are covalently attached to the linker usingconventional chemical techniques providing for covalent linkage of thePEG group to the linker. The linker, in turn, may be covalently attachedto the otherwise, non-PEG substituted compounds of the presentinvention. Reaction chemistries resulting in such linkages are wellknown in the art. Such reaction chemistries involve the use ofcomplementary functional groups on the linker, the non-PEG substitutedcompound of the present invention and the PEG groups. Preferably, thecomplementary functional groups on the linker are selected relative tothe functional groups available on the PEG group for bonding or whichcan be introduced onto the PEG group for bonding. Again, suchcomplementary functional groups are well known in the art.

Such polymers have a number average molecular weight of from about 100to 100,000; preferably from about 1,000 to 50,000; more preferably fromabout 10,000 to about 40,000.

The polymer conjugates of the invention may provide enhanced in vivoretention as compared to the non-conjugated compounds. The improvedretention of the conjugate within the body results in lower requireddosages of the drug, which in turn results in fewer side effects andreduced likelihood of toxicity. In addition, the drug formulationcomprising these polymer conjugates may be administered less frequentlyto the patient while achieving a similar or improved therapeutic effect.The conjugates of this invention have improved inhibition, in vivo, ofadhesion of leukocytes to endothelial cells mediated by VLA-4 bycompetitive binding to VLA-4. Preferably, the compounds of thisinvention can be used in I.V. formulations.

The therapeutic dosage of the polymer conjugates of the presentinvention will vary according to, for example, the particular use forwhich the treatment is made, the manner of administration of thecompound, the health and condition of the patient, and the judgment ofthe prescribing physician. For example, for intravenous administration,the dose will typically be in the range of about 20 μg to about 2000 μgper kilogram body weight, preferably about 20 μg to about 500 μg, morepreferably about 100 μg to about 300 μg per kilogram body weight.Suitable dosage ranges for intranasal administration are generally about0.1 pg to I mg per kilogram body weight. Effective doses can beextrapolated from dose-response curves derived from in vitro or animalmodel test systems.

When formulated and administered as polymer conjugates, the compounds orconjugates of this invention are characterized as containing one or morepolyethylene glycol substituents covalently attached thereto. Withoutbeing limited to any theory, the improved serum half-life is believed tobe associated with covalent conjugation of at least one polyethyleneglycol entity onto the structure of the compound.

Accordingly, the compounds of the present invention may be PEGderivatives of formula XXII below:

-   -   wherein    -   R is selected from the group consisting of a PEG moiety, amino,        substituted amino, alkyl and substituted alkyl wherein each        amino, substituted amino, alkyl and substituted alkyl is        optionally substituted with a PEG moiety wherein, in each case,        the PEG moiety optionally comprises a linker which covalently        links the PEG moiety;    -   Ar¹ is selected from the group consisting of aryl, substituted        aryl, heteroaryl and substituted heteroaryl wherein each of        aryl, substituted aryl, heteroaryl and substituted heteroaryl is        optionally substituted with a PEG moiety wherein the PEG moiety        optionally comprises a linker which covalently links the PEG        moiety to Ar¹;    -   Ar² is selected from the group consisting of aryl, substituted        aryl, heteroaryl and substituted heteroaryl wherein each of        aryl, substituted aryl, heteroaryl and substituted heteroaryl is        optionally substituted with a PEG moiety wherein the PEG moiety        optionally comprises a linker which covalently links the PEG        moiety to Ar²;    -   X is selected from the group consisting of —S—, —SO—, —SO₂ and        optionally substituted —CH₂—;    -   Y is selected from the group consisting of —O— and —NR¹— wherein        R¹ is selected from the group consisting of hydrogen and alkyl;    -   W is selected from the group consisting of a PEG moiety which        optionally comprises a linker and —NR²R³ wherein R² and R³ are        independently selected from the group consisting of alkyl,        substituted alkyl, and where R² and R³, together with the        nitrogen atom bound thereto, form a heterocyclic ring or a        substituted heterocyclic ring wherein each of alkyl, substituted        alkyl, heterocyclic and substituted heterocyclic is optionally        substituted with a PEG moiety which further optionally comprises        a linker;    -   m is an integer equal to 0, 1 or 2;    -   n is an integer equal to 0 to 2; and    -   pharmaceutically acceptable salts thereof;    -   provided that at least one of R, Ar¹, Ar², W and —NR²R³ contains        a PEG moiety;    -   further provided that when R is a PEG moiety, n is one and X is        not —S—, —SO—, or —SO₂—;    -   and still further provided that the compound of formula XXII has        a molecular weight of no more than 100,000.

Preferably the PEG derivates of formula XXII are the of the L isomer asshown below:

In another aspect, the compounds of the present invention may be PEGderivatives of formula XXIII below:

-   -   wherein    -   Ar¹, Ar², Y and W are as defined above; and    -   pharmaceutically acceptable salts thereof;    -   provided that at least one of Ar¹, Ar², W and —NR²R³ contains a        PEG moiety which optionally comprises a linker;    -   and further provided that the compound of formula XXIII has a        molecular weight of no more than 100,000.

In another aspect, the compounds of the present invention may be PEGderivatives of formula XXIV below:

-   -   wherein    -   R, Ar¹, Ar², Y, W and n are as defined above; and    -   pharmaceutically acceptable salts thereof;    -   provided that at least one of R, Ar¹, Ar², W and —NR²R³ contains        a PEG moiety which optionally comprises a linker;    -   and further provided that the compound of formula XXVI has a        molecular weight of no more than 100,000.

In another aspect, the compounds of the present invention may be PEGderivatives of formula XXV below:

-   -   wherein    -   R, R², R³, Ar¹, Ar² and n are as defined above; and    -   pharmaceutically acceptable salts thereof;    -   provided that at least one of R, Ar¹, Ar², and —NR²R³ contains a        PEG moiety which optionally comprises a linker;    -   and further provided that the compound of formula XXV has a        molecular weight of no more than 100,000.

In another of its aspects, the compound of this invention is directed toa PEG derivative of formula XXVI below:

-   -   wherein    -   R², R³, Ar¹, and Ar² are as defined above; and    -   pharmaceutically acceptable salts thereof;    -   provided that at least one of Ar¹, Ar² and —NR²R³ contains a PEG        moiety which optionally comprises a linker;    -   and further provided that the compound of formula XXVI has a        molecular weight of no more than 100,000.

In another aspect, the compounds of this invention can be PEGderivatives of formula XXVII:

-   -   wherein    -   R⁴ is a PEG moiety which optionally comprises a linker;    -   R⁵ is selected from the group consisting of alkyl and        substituted alkyl;    -   Ar³ is selected from the group consisting of aryl, substituted        aryl, heteroaryl and substituted heteroaryl;    -   X is selected from the group consisting of —S—, —SO—, and —SO₂—        or optionally substituted —CH₂—;    -   m is an integer equal to 0, 1 or 2;    -   n is an integer equal to 0 to 2; and    -   pharmaceutically acceptable salts thereof;    -   provided that the compound of formula XXVII has a molecular        weight of no more than 100,000.

In another aspect, the compound of the invention can be a PEG derivativeof formula XXVIII:

-   -   wherein    -   R⁴ is a PEG moiety which optionally comprises a linker;    -   R⁵ is selected from the group consisting of alkyl and        substituted alkyl;    -   Ar³ is selected from the group consisting of aryl, substituted        aryl, heteroaryl and substituted heteroaryl;    -   n is an integer equal to 0 to 2; and    -   pharmaceutically acceptable salts thereof;    -   provided that the compound of formula XXVIII has a molecular        weight of no more than 100,000.

In another aspect, the compound of the invention can be a PEG derivativeof formula XXIX:

-   -   wherein    -   R⁴ is a PEG moiety which optionally comprises a linker;    -   Ar³ is selected from the group consisting of aryl, substituted        aryl, heteroaryl and substituted heteroaryl;    -   pharmaceutically acceptable salts thereof;    -   provided that the compound of formula XXIX has a molecular        weight of no more than 100,000.

Preferably, when Ar¹ does not contain a PEG moiety, Ar¹ in formulasXXII-XXVI and Ar³ in formulas XXVII-XXIX is selected from the groupconsisting of:

-   -   phenyl,    -   4-methylphenyl,    -   4-t-butylphenyl,    -   2,4,6-trimethylphenyl,    -   2-fluorophenyl,    -   3-fluorophenyl,    -   4-fluorophenyl,    -   2,4-difluorophenyl,    -   3,4-difluorophenyl,    -   3,5-difluorophenyl,    -   2-chlorophenyl,    -   3-chlorophenyl,    -   4-chlorophenyl,    -   3,4-dichlorophenyl,    -   3,5-dichlorophenyl,    -   3-chloro-4-fluorophenyl,    -   4-bromophenyl,    -   2-methoxyphenyl,    -   3-methoxyphenyl,    -   4-methoxyphenyl,    -   3,4-dimethoxyphenyl,    -   4-t-butoxyphenyl,    -   4-(3′-dimethylamino-n-propoxy)-phenyl,    -   2-carboxyphenyl,    -   2-(methoxycarbonyl)phenyl,    -   4-(H₂NC(O)—)phenyl,    -   4-(H₂NC(S)—)phenyl,    -   4-cyanophenyl,    -   4-trifluoromethylphenyl,    -   4-trifluoromethoxyphenyl,    -   3,5-di-(trifluoromethyl)phenyl,    -   4-nitrophenyl,    -   4-aminophenyl,    -   4-(CH₃C(O)NH—)phenyl,    -   4-(PhNHC(O)NH—)phenyl,    -   4-amidinophenyl,    -   4-methylamidinophenyl,    -   4-[CH₃SC(═NH)-]phenyl,    -   4-chloro-3-[H₂NS(O)₂-]phenyl,    -   1-naphthyl,    -   2-naphthyl,    -   pyridin-2-yl,    -   pyridin-3-yl,    -   pyridine-4-yl,    -   pyrimidin-2-yl,    -   quinolin-8-yl,    -   2-(trifluoroacetyl)-1,2,3,4-tetrahydroisoquinolin-7-yl,    -   2-thienyl,    -   5-chloro-2-thienyl,    -   2,5-dichloro-4-thienyl,    -   1-N-methylimidazol-4-yl,    -   1-N-methylpyrazol-3-yl,    -   1-N-methylpyrazol-4-yl,    -   1-N-butylpyrazol-4-yl,    -   1-N-methyl-3-methyl-5-chloropyrazol-4-yl,    -   1-N-methyl-5-methyl-3-chloropyrazol-4-yl,    -   2-thiazolyl, and    -   5-methyl-1,3,4-thiadiazol-2-yl.

When Ar¹ contains a PEG group, Ar¹ is preferably of the formula:—Ar¹—Z—(CH₂CHR⁷O)_(p)R⁸

-   -   wherein    -   Ar¹ is selected from the group consisting of aryl, substituted        aryl, heteroaryl, and substituted heteroaryl,    -   Z is selected from the group consisting of a covalent bond, a        linking group of from 1 to 40 atoms, —O—, and —NR⁹—, where R⁹ is        selected from the group consisting of hydrogen and alkyl,    -   R⁷ is selected from the group consisting of hydrogen and methyl;    -   R⁸ is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, aryl, substituted aryl, and —CH₂CHR⁷NR¹⁰R¹¹        where R⁷ is as defined above and R¹⁰ and R¹¹ are independently        selected from the group consisting of hydrogen and alkyl; and    -   p is an integer such that the molecular weight of the PEG moiety        ranges from 100 to 100,000.

Preferably, when R does not contain a PEG moiety, the substituent of theformula:

-   -   where X, m and n are as defined above, and R′ is alkyl or        substituted alkyl is preferably selected from the group        consisting of:    -   azetidinyl, thiazolidinyl, piperidinyl, piperazinyl,        thiomorpholinyl pyrrolidinyl, 4-hydroxypyrrolidinyl,        4-oxopyrrolidinyl, 4-fluoropyrrolidinyl,        4,4-difluoropyrrolidinyl,        4-(thiomorpholin-4-ylC(O)O-)pyrrolidinyl,        4-[CH₃S(O)₂O-]pyrrolidinyl, 3-phenylpyrrolidinyl,        3-thiophenylpyrrolidinyl, 4-aminopyrrolidinyl,        3-methoxypyrrolidinyl, 4,4-dimethylpyrrolidinyl,        4-N-Cbz-piperazinyl, 4-[CH₃S(O)₂-]piperazinyl,        5,5-dimethylthiazolindin-4-yl, 1,1-dioxo-thiazolidinyl,        1,1-dioxo-5,5-dimethylthiazolidin-2-yl and        1,1-dioxothiomorpholinyl.

When the substituent of the formula:

-   -   contains a PEG moiety, then preferably the substituent is of the        formula:    -   wherein    -   m is an integer equal to zero, one or two;    -   Z is selected from the group consisting of a covalent bond, a        linking group of from 1 to 40 atoms, —O—, and —NR⁹—, where R⁹ is        selected from the group consisting of hydrogen and alkyl,    -   R⁷ is selected from the group consisting of hydrogen and methyl;    -   R⁸ is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, aryl, substituted aryl, and —CH₂CHR⁷NR¹⁰R¹¹        where R⁷ is as defined above and R¹⁰ and R¹¹ are independently        selected from the group consisting of hydrogen and alkyl; and    -   p is an integer such that the molecular weight of the PEG moiety        ranges from 100 to 100,000.

Preferably, when Ar² does not contain a PEG moiety, Ar² in formulas I-Vis preferably selected from the group consisting of phenyl, 2-pyridyl,3-pyridyl, 4-pyridyl, and 4-pyrid-2-onyl.

When Ar² contains a PEG moiety, Ar² in formulas XXII-XXVI is preferablyrepresented by the formula:

-   -   where Ar² is selected from the group consisting of aryl,        substituted aryl, heteroaryl and substituted heteroaryl;    -   Z is selected from the group consisting of a covalent bond, a        linking group of from 1 to 40 atoms, —O—, and —NR⁹—, where R⁹ is        selected from the group consisting of hydrogen and alkyl,    -   R⁷ is selected from the group consisting of hydrogen and methyl;    -   R⁸ is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, aryl, substituted aryl, and —CH₂CHR⁷NR¹⁰R¹¹        where R⁷ is as defined above and R¹⁰ and R¹¹ are independently        selected from the group consisting of hydrogen and alkyl; and    -   p is an integer such that the molecular weight of the PEG moiety        ranges from 100 to 100,000.

Preferably, in formulas XXII-XXIV, —YC(O)W is —OC(O)NR²R³. When R² andR³ do not contain a PEG moiety, —OC(O)NR²R³ in formulas XXII-XXVI ispreferably selected from the group:

-   -   (CH₃)₂NC(O)O—,    -   (piperidin-1-yl)C(O)O—,    -   (4-hydroxypiperidin-1-yl)C(O)O—,    -   (4-formyloxypiperidin-1-yl)C(O)O—,    -   (4-ethoxycarbonylpiperidin-1-yl)C(O)O—,    -   (4-carboxylpiperidin-1-yl)C(O)O—,    -   (3-hydroxymethylpiperidin-1-yl)C(O)O—,    -   (4-hydroxymethylpiperidin-1-yl)C(O)O—,    -   (4-piperidon-1-yl ethylene ketal)C(O)O—,    -   (piperazin-1-yl)-C(O)O—,    -   (1-Boc-piperazin-4-yl)-C(O)O—,    -   (4-methylpiperazin-1-yl)C(O)O—,    -   (4-methylhomopiperazin-1-yl)C(O)O—,    -   (4-(2-hydroxyethyl)piperazin-1-yl)C(O)O—,    -   (4-phenylpiperazin-1-yl)C(O)O—,    -   (4-(pyridin-2-yl)piperazin-1]-yl)C(O)O—,    -   (4-(4-trifluoromethylpyridin-2-yl)piperazin-1-yl)C(O)O—,    -   (4-(pyrimidin-2-yl)piperazin-1-yl)C(O)O—,    -   (4-acetylpiperazin-1-yl)C(O)O—,    -   (4-(phenylC(O)-)piperazin-1-yl)C(O)O—,    -   (4-(pyridin-4′-ylC(O)-)piperazin-1-yl)C(O)O,    -   (4-(phenylNHC(O)-)piperazin-1-yl)C(O)O—,    -   (4-(phenylNHC(S)-)piperazin-1-yl)C(O)O—,    -   (4-methanesulfonylpiperazin-1-yl-C(O)O—,    -   (4-trifluoromethanesulfonylpiperazin-1-yl-C(O)O—,    -   (morpholin-4-yl)C(O)O—,    -   (thiomorpholin-4-yl)C(O)O—,    -   (thiomorpholin-4′-yl sulfone)-C(O)O—,    -   (pyrrolidin-1-yl)C(O)O—,    -   (2-methylpyrrolidin-1-yl)C(O)O—,    -   (2-(methoxycarbonyl)pyrrolidin-1-yl)C(O)O—,    -   (2-(hydroxymethyl)pyrrolidin-1-yl)C(O)O—,    -   (2-(N,N-dimethylamino)ethyl)(CH₃)NC(O)O—,    -   (2-(N-methyl-N-toluene-4-sulfonylamino)ethyl)(CH₃)N—C(O)O—,    -   (2-(morpholin-4-yl)ethyl)(CH₃)NC(O)O—,    -   (2-(hydroxy)ethyl)(CH₃)NC(O)O—,    -   bis(2-(hydroxy)ethyl-)NC(O)O—,    -   (2-(formyloxy)ethyl)(CH₃)NC(O)O—,    -   (CH₃OC(O)CH₂)HNC(O)O—, and    -   2-[(phenylNHC(O)O-)ethyl-]HNC(O)O—.

When R² and/or R³ comprise a PEG moiety, the PEG moiety is preferablyrepresented by the formula:—Z′—(CH₂CHR⁷O)_(p)R⁸

-   -   Z′ is selected from the group consisting of a covalent bond and        a linking group of from 1 to 40 atoms;.    -   R⁷ is selected from the group consisting of hydrogen and methyl;    -   R⁸ is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, aryl, substituted aryl, and —CH₂CHR⁷NR¹⁰R¹¹        where R⁷ is as defined above and R¹⁰ and R¹¹ are independently        selected from the group consisting of hydrogen and alkyl; and    -   p is an integer such that the molecular weight of the PEG moiety        ranges from 100 to 100,000.

Preferred —YC(O)W substituents comprising a PEG moiety include thefollowing:

-   -   where φ or C₆H₅ is phenyl and p is an integer such that the        molecular weight of the PEG moiety ranges from about 100 to        100,000 and v is 1 to 5.

Preferred PEG derivatives of this invention include those set forthbelow:

where, in each case, PEG is a methyl capped polyoxyethylene group havinga molecular weight (Mw) of approximately 20,000.

“Linking group” or “linker” of from 1 to 40 atoms is a di- to hexavalentgroup or groups that covalently links a non-PEG substituted compound offormula I (i.e., none of Ar¹, Ar², R or —Y—C(O)—W— contain a PEG group)with 1 to 5 PEG groups. Each linker may be chiral or achiral, linear,branched or cyclic and may be homogenous or heterogeneous in its atomcontent (e.g., linkers containing only carbon atoms or linkerscontaining carbon atoms as well as one or more heteroatoms present onthe linker in the form of alcohols, ketones, aldehydes, carboxyl groups,amines, amides, carbamates, ureas, thiols, ethers, etc., or residuesthereof) Preferably, the linker contains 1 to 25 carbon atoms and 0 to15 heteroatoms selected from oxygen, NR²², sulfur, —S(O)— and —S(O)₂—,where R²² is as defined above.

The PEG group or groups are covalently attached to the linker usingconventional chemical techniques providing for covalent linkage of thePEG group to the linker. The linker, in turn, is covalently attached tothe otherwise, non-PEG substituted compound of formula I. Reactionchemistries resulting in such linkages are well known in the art. Suchreaction chemistries involve the use of complementary functional groupson the linker, the non-PEG substituted compound of formula XXII and thePEG groups. Preferably, the complementary functional groups on thelinker are selected relative to the functional groups available on thePEG group for bonding or which can be introduced onto the PEG group forbonding. Again, such complementary functional groups are well known inthe art. For example, reaction between a carboxylic acid of either thelinker or the PEG group and a primary or secondary amine of the PEGgroup or the linker in the presence of suitable, well-known activatingagents results in formation of an amide bond covalently linking the PEGgroup to the linker; reaction between an amine group of either thelinker or the PEG group and a sulfonyl halide of the PEG group or thelinker results in formation of a sulfonamide bond covalently linking thePEG group to the linker; and reaction between an alcohol or phenol groupof either the linker or the PEG group and an alkyl or aryl halide of thePEG group or the linker results in formation of an ether bond covalentlylinking the PEG group to the linker.

Table 9 below illustrates numerous complementary reactive groups and theresulting bonds formed by reaction therebetween. TABLE 9 RepresentativeComplementary Binding Chemistries First Reactive Group Second ReactiveGroup Linkage Hydroxyl Isocyanate urethane Amine Epoxide β-hydroxyaminesulfonyl halid Amine sulfonamide Carboxyl Amine amide Hydroxylalkyl/aryl halide ether

Preferred linkers include, by way of example, the following —O—, —NR²²—,—NR²²C(O)O—, —OC(O)NR²², —NR²²C(O)—, —C(O)NR²²—, —NR²²C(O)NR²²—,-alkylene-NR²²C(O)O—, -alkylene-NR²²C(O)NR²²—, -alkylene-OC(O)NR²²,-alkylene-NR²²—, -alkylene-O—, -alkylene-NR³C(O)—, -alkylene-C(O)NR²²—,—NR²²C(O)O-alkylene-, —NR²²C(O)NR²²-alkylene-, —OC(O)NR²²-alkylene,—NR²²-alkylene-, —O-alkylene-, —NR²²C(O)-alkylene-, —C(O)NR²²-alkylene-,-alkylene-NR²²C(O)O-alkylene-, -alkylene-NR³C(O)NR²²-alkylene-,-alkylene-OC(O)NR²²-alkylene-, -alkylene-NR²²-alkylene-,alkylene-O-alkylene-, -alkylene-NR²²C(O)-alkylene-, —C(O)NR²²-alkylene-,and

-   -   is selected from the group consisting of aryl, substituted aryl,        cycloalkyl, substituted cycloalkyl, heteroaryl, substituted        heteroaryl, heterocyclic and substituted heterocyclic, and B and        C are independently selected from the group consisting of a        bond, —O—, CO, —NR²²—, —NR²²C(O)O—, —OC(O)NR²²—, —NR²²C(O)—,        —C(O)NR²²—, —NR²²C(O)NR²²—, -alkylene-NR²²C(O)O—,        -alkylene-NR²²C(O)NR²²—, -alkylene-OC(O)NR²²—, -alkylene-NR²²—,        -alkylene-O—, -alkylene-NR²²C(O)—, alkylene-C(O)NR²²—,        —NR²²C(O)O-alkylene-, —NR²²C(O)NR²²-alkylene-,        —OC(O)NR²²-alkylene-, —NR²²-alkylene-, —O-alkylene-, —NR²²        C(O)-alkylene-, —C(O)NR²²-alkylene-,        -alkylene-NR²²C(O)O-alkylene-, -alkylene-NR²²C(O)NR²²-alkylene-,        -alkylene-OC(O)NR²²-alkylene-, -alkylene-NR²²-alkylene-,        alkylene-O-alkylene-, -alkylene-NR²²C(O)-alkylene-, and —C(O)NR        22-alkylene-, where R²² is as defined above.

“PEG” or “PEG moiety” refers to polymers comprising multiple oxyalkyleneunits. Such polymers are optionally mono-capped with a substituentpreferably selected from alkyl, aryl, substituted alkyl, and substitutedaryl. Inclusive of such polymers are those diamino cappedpolyoxyalkylene polymers which are known in the art as Jeffamines®.Still further, such polymers can optionally contain one or morenon-oxyalkylene units such as the commercially availablepoly[di(ethylene glycol)adipates, poly[di(ethylene glycol)phthalatediols, and the like. Also included are block copolymers of oxyalkylene,polyethylene glycol, polypropylene glycol, and polyoxyethylenated polyolunits.

Such polymers have a number average molecular weight of from about 100to 100,000; preferably from about 1,000 to 50,000; more preferably fromabout 10,000 to about 40,000. In a particularly preferred embodiment,the molecular weight of the total amount of PEG arising from single ormultiple PEG moieties bound in the molecule does not exceed 100,000;more preferably 50,000 and even more preferably 40,000.

In a preferred embodiment, the -[linking group]_(u)-PEG group where u iszero or one can be represented by the formula:-Z′-[(CH₂CHR⁷O)_(p)R⁸]_(t)

-   -   where Z′ is selected from the group consisting of a covalent        bond, a linking group of from 1 to 40 atoms, —O—, —S—, —NR²²—,        —C(O)O—, —C(O)NR²²—, and —C(O)— where R²² is selected from the        group consisting of hydrogen and alkyl,    -   R⁷ is selected from the group consisting of hydrogen and methyl;    -   R⁸ is selected from the group consisting of hydrogen, alkyl,        substituted alkyl, aryl, substituted aryl, —CH₂CHR⁷SR⁷ and        -CH₂CHR⁷NR¹⁰R¹¹ where R⁷ is as defined above and R¹⁰ and R¹¹ are        independently selected from the group consisting of hydrogen and        alkyl;    -   p is an integer such that the molecular weight of the PEG moiety        ranges from 100 to 100,000; and    -   t is an integer from 1 to 5 provided that t is one less than the        valency of the linking group and is one when there is no linking        group.

When Z′ is linking group, multiple PEG groups can be present. Forexample, if the linking group is trivalent, then 2 PEG groups can beattached and the remaining valency is employed to link to the moleculeof formula XXII. Preferably the number of PEG groups is 1 or 2. In anyevent, when multiple PEG groups are present, the total aggregatemolecular weight of the PEG groups does not exceed 100,000.

PEG Derivative Preparation

The compounds of this invention can be prepared from readily availablestarting materials using the following general methods and procedures.It will be appreciated that where typical or preferred processconditions (i.e., reaction temperatures, times, mole ratios ofreactants, solvents, pressures, etc.) are given, other processconditions can also be used unless otherwise stated. Optimum reactionconditions may vary with the particular reactants or solvent used, butsuch conditions can be determined by one skilled in the art by routineoptimization procedures.

Additionally, as will be apparent to those skilled in the art,conventional protecting groups may be necessary to prevent certainfunctional groups from undergoing undesired reactions. Suitableprotecting groups for various functional groups as well as suitableconditions for protecting and deprotecting particular functional groupsare well known in the art. For example, numerous protecting groups aredescribed in T. W. Greene and G. M. Wuts, Protecting Groups in OrganicSynthesis, Second Edition, Wiley, N.Y., 1991, and references citedtherein.

Furthermore, the compounds of this invention will typically contain oneor more chiral centers. Accordingly, if desired, such compounds can beprepared or isolated as pure stereoisomers, i.e., as individualenantiomers or diastereomers, or as stereoisomer-enriched mixtures. Allsuch stereoisomers (and enriched mixtures) are included within the scopeof this invention, unless otherwise indicated. Pure stereoisomers (orenriched mixtures) may be prepared using, for example, optically activestarting materials or stereoselective reagents well-known in the art.Alternatively, racemic mixtures of such compounds can be separatedusing, for example, chiral column chromatography, chiral resolvingagents and the like.

The compounds of this invention are preferably characterized bycontaining one or more PEG moieties at one of several sites of acompound of formula XXIIa:

Specifically, the PEG moiety can be incorporated into the Ar¹substituent, the R substituent, the Ar² substituent and/or in the—YC(O)W substituent wherein the PEG moiety is either directly attachedor is attached via a linker. The synthetic protocol for insertion of aPEG moiety at each of these positions is similar and entails reaction ofa functional group on the PEG moiety or the linking group covalentlybound to the PEG moiety with a complementary functional group on thenon-PEG substituted compounds of formula XXIIa.

Initially, non-PEG substituted compounds of formula XXIIa are well knownin the art and are exemplified in a number of issued patents including,without limitation, U.S. Pat. Nos. 6,489,300 and 6,436,904 both of whichare incorporated herein by reference in their entirety. Non-PEG variantsof compounds of formula Ia include those having complementary functionalgroups or groups derivatizable to complementary functional groups on oneor more of the Ar¹, R, Ar² and —YC(O)W moieties. For illustrativepurposes, compounds having a complementary functional group (—OH) on theAr² moiety (e.g., tyrosine) are recited below as a suitable startingpoint for addition of a PEG group to the molecule either directly orthrough a linker.

Such compounds can be prepared by first coupling a heterocyclic aminoacid, 1, with an appropriate aryl sulfonyl chloride as illustrated inScheme 1 below:

where R, Ar¹, X, m and n are as defined above.

Specifically, in Scheme 1 above, heterocyclic amino acid, 1, is combinedwith a stoichiometric equivalent or excess amount (preferably from about1.1 to about 2 equivalents) of arylsulfonyl halide, 2, in a suitableinert diluent such as dichloromethane and the like. Generally, thereaction is conducted at a temperature ranging from about −70° C. toabout 40° C. until the reaction is substantially complete, whichtypically occurs within 1 to 24 hours. Preferably, the reaction isconducted in the presence of a suitable base to scavenge the acidgenerated during the reaction. Suitable bases include, by way ofexample, tertiary amines, such as triethylamine, diisopropylethylamine,N-methyl-morpholine and the like. Alternatively, the reaction can beconducted under Schotten-Baumann-type conditions using an aqueous alkalisolution such as an aqueous solution of sodium hydroxide, an aqueousphosphate solution buffered to pH 7.4, and the like. The resultingproduct, 3, can be recovered by conventional methods, such aschromatography, filtration, evaporation, crystallization, and the likeor, alternatively, used in the next step without purification and/orisolation.

Heterocyclic amino acids, 1, employed in the above reaction are eitherknown compounds or compounds that can be prepared from known compoundsby conventional synthetic procedures. Examples of suitable amino acidsfor use in this reaction include, but are not limited to, L-proline,trans-4-hydroxyl-L-proline, cis-4-hydroxyl-L-proline,trans-3-phenyl-L-proline, cis-3-phenyl-L-proline, L-(2-methyl)proline,L-pipecolinic acid, L-azetidine-2-carboxylic acid,L-thiazolidine-4-carboxylic acid,L-(5,5-dimethyl)thiazolidine-4-carboxylic acid,L-thiamorpholine-3-carboxylic acid. If desired, the correspondingcarboxylic acid esters of the amino acids, 1, such as the methyl esters,ethyl esters, t-butyl esters, and the like, can be employed in the abovereaction with the arylsulfonyl chloride. Subsequent hydrolysis of theester group to the carboxylic acid using conventional reagents andconditions, i.e., treatment with an alkali metal hydroxide in an inertdiluent such as methanol/water, then provides the N-sulfonyl amino acid,3.

Similarly, the arylsulfonyl chlorides, 2, employed in the above reactionare either known compounds or compounds that can be prepared from knowncompounds by conventional synthetic procedures. Such compounds aretypically prepared from the corresponding sulfonic acid, i.e., fromcompounds of the formula Ar¹SO₃H where Ar¹ is as defined above, usingphosphorous trichloride and phosphorous pentachloride. This reaction isgenerally conducted by contacting the sulfonic acid with about 2 to 5molar equivalents of phosphorous trichloride and phosphorouspentachloride, either neat or in an inert solvent, such asdichloromethane, at temperature in the range of about 0° C. to about 80°C. for about 1 to about 48 hours to afford the sulfonyl chloride.Alternatively, the arylsulfonyl chlorides, 2, can be prepared from thecorresponding thiol compound, i.e., from compounds of the Ar¹—SH whereAr¹ is as defined herein, by treating the thiol with chlorine (Cl₂) andwater under conventional reaction conditions.

Alternatively, arylsulfonyl chlorides, 2, employed in the above reactionmay be prepared by chlorosulfonylation of substituted benzene orheterocycloalkyl group using Cl—SO₃H.

Examples of arylsulfonyl chlorides suitable for use in this inventioninclude, but are not limited to, benzenesulfonyl chloride,1-naphthalenesulfonyl chloride, 2-naphthalenesulfonyl chloride,p-toluenesulfonyl chloride, o-toluenesulfonyl chloride,4-acetamidobenzenesulfonyl chloride, 4-tert-butylbenzenesulfonylchloride, 4-bromobenzenesulfonyl chloride, 2-carboxybenzenesulfonylchloride, 4-cyanobenzenesulfonyl chloride, 3,4-dichlorobenzenesulfonylchloride, 3,5-dichlorobenzenesulfonyl chloride,3,4-dimethoxybenzenesulfonyl chloride,3,5-ditrifluoromethylbenzenesulfonyl chloride, 4-fluorobenzenesulfonylchloride, 4-methoxybenzenesulfonyl chloride,2-methoxycarbonylbenzenesulfonyl chloride, 4-methylamido-benzenesulfonylchloride, 4-nitrobenzenesulfonyl chloride,4-trifluoromethyl-benzenesulfonyl chloride,4-trifluoromethoxybenzenesulfonyl chloride,2,4,6-trimethylbenzenesulfonyl chloride, 2-thiophenesulfonyl chloride,5-chloro-2-thiophenesulfonyl chloride, 2,5-dichloro-4-thiophenesulfonylchloride, 2-thiazolesulfonyl chloride, 2-methyl-4-thiazolesulfonylchloride, 1-methyl-4-imidazolesulfonyl chloride,1-methyl-4-pyrazolesulfonyl chloride,5-chloro-1,3-dimethyl-4-pyrazolesulfonyl chloride, 3-pyridinesulfonylchloride, 2-pyrimidinesulfonyl chloride and the like. If desired, asulfonyl fluoride, sulfonyl bromide or sulfonic acid anhydride may beused in place of the sulfonyl chloride in the above reaction to form theN-sulfonyl amino acid, 3.

The N-arylsulfonyl amino acid, 3, is then coupled to commerciallyavailable tyrosine esters as shown in Scheme 2 below:

where R, Ar¹, X, m and n are as defined above, R^(a) is hydrogen oralkyl but preferably is an alkyl group such as t-butyl, Z representsoptional substitution on the aryl ring and q is zero, one or two.

This coupling reaction is typically conducted using well-known couplingreagents such as carbodiimides, BOP reagent(benzotriazol-1-yloxy-tris(dimethylamino)-phosphoniumhexafluorophosphonate) and the like. Suitable carbodiimides include, byway of example, dicyclohexylcarbodiimide (DCC),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) and the like. Ifdesired, polymer supported forms of carbodiimide coupling reagents mayalso be used including, for example, those described in TetrahedronLetters, 34(48), 7685 (1993). Additionally, well-known couplingpromoters, such as N-hydroxysuccinimide, 1-hydroxybenzotriazole and thelike, may be used to facilitate the coupling reaction.

This coupling reaction is typically conducted by contacting theN-sulfonylamino acid, 3, with about 1 to about 2 equivalents of thecoupling reagent and at least one equivalent, preferably about 1 toabout 1.2 equivalents, of tyrosine derivative, 4, in an inert diluent,such as dichloromethane, chloroform, acetonitrile, tetrahydrofuran,N,N-dimethylformamide and the like. Generally, this reaction isconducted at a temperature ranging from about 0° C. to about 37° C. forabout 12 to about 24 hours. Upon completion of the reaction, thecompound 5 is recovered by conventional methods includingneutralization, evaporation, extraction, precipitation, chromatography,filtration, and the like.

Alternatively, the N-sulfonyl amino acid, 3 can be converted into anacid halide which is then coupled with compound, 4, to provide compound5. The acid halide can be prepared by contacting compound 3 with aninorganic acid halide, such as thionyl chloride, phosphoroustrichloride, phosphorous tribromide or phosphorous pentachloride, orpreferably, with oxalyl chloride under conventional conditions.Generally, this reaction is conducted using about 1 to 5 molarequivalents of the inorganic acid halide or oxalyl chloride, either neator in an inert solvent, such as dichloromethane or carbon tetrachloride,at temperature in the range of about 0° C. to about 80° C. for about 1to about 48 hours. A catalyst, such as DMF, may also be used in thisreaction.

The acid halide of N-sulfonyl amino acid, 3, is then contacted with atleast one equivalent, preferably about 1.1 to about 1.5 equivalents, ofthe tyrosine derivative, 4, in an inert diluent, such asdichloromethane, at a temperature ranging from about −70° C. to about40° C. for about 1 to about 24 hours. Preferably, this reaction isconducted in the presence of a suitable base to scavenge the acidgenerated during the reaction. Suitable bases include, by way ofexample, tertiary amines, such as triethylamine, diisopropylethylamine,N-methylmorpholine and the like. Alternatively, the reaction can beconducted under Schotten-Baumann-type conditions using aqueous alkali,such as sodium hydroxide and the like. Upon completion of the reaction,compound 5 is recovered by conventional methods includingneutralization, evaporation, extraction, precipitation, chromatography,filtration, and the like.

Aternatively, compound 5 can be prepared by first forming a diamino acidderivative and then coupling the diamino acid to the arylsulfonylhalide, 2, as shown in scheme 3 below:

where R, R^(a), Ar¹, X, Z, m, n and q are as defined above.

The diamino acid, 6, can be readily prepared by coupling amino acid, 1,with amino acid, 4, using conventional amino acid coupling techniquesand reagents, such carbodiimides, BOP reagent and the like, as describedabove. Diamino acid, 6, can then be sulfonated using sulfonyl chloride,2, and using the synthetic procedures described above to providecompound 7.

The tyrosine derivatives, 4, employed in the above reactions are eitherknown compounds or compounds that can be prepared from known compoundsby conventional synthetic procedures. For example, tyrosine derivatives,4, suitable for use in the above reactions include, but are not limitedto, L-tyrosine methyl ester, L-tyrosine t-butyl-esterL-3,5-diiodotyrosine methyl ester, L-3-iodotyrosine methyl ester,β-(4-hydroxy-naphth-1-yl)-L-alanine methyl ester,β-(6-hydroxy-naphth-2-yl)-L-alanine methyl ester, and the like. Ifdesired, of course, other esters or amides of the above-describedcompounds may also be employed.

The N-arylsulfonyl-heterocyclic amino acid-tyrosine derivative, 7, canbe used as a starting point to prepare PEG derivatives at the Ar² groupby coupling reactions shown in Schemes 4-14 below which couplingreactions are illustrative only in demonstrating how PEG moieties can beintroduced. In some cases, the PEG moiety can be directly introducedonto the phenoxy group and, in other cases, the PEG moiety can beintroduced by linkage through a linker moiety.

Specifically, Scheme 4 illustrates the following:

wherein Ar¹, R, R^(a), m, n, q, X, and Z are as defined above whereas Qis oxygen, sulfur and NH, Pg is an amine protecting group such as CBZ,Boc, etc, which is preferably orthogonally removeable as compared to theR^(a) carboxyl protecting group and PEG is preferably a methyl cappedpoly(oxyethylene) group having a molecular weight of from 100 to100,000.

In Scheme 4, the PEG moiety is covalently attached to theN-piperazinylcarbonyltyrosine moiety (R²/R³ are joined together with thenitrogen atom attached thereto to form a piperazine ring) via a linkerentity which constitutes the group:

Specifically, in Scheme 4, compound 7, prepared as above, is combinedwith at least an equivalent and preferably an excess of 4-nitrophenylchloroformate, 8, in a suitable solvent such as methylene chloride,chloroform and the like and preferably under an inert atmosphere. Thereaction is preferably conducted at a temperature of from about −40° toabout 0° C. in the presence of a suitable base to scavenge the acidgenerated. Suitable bases include, by way of example, triethylamine,diisopropylethylamine, and the like. After formation of the intermediatemixed carbonate (not shown), at least an approximately equimolar amountof N-Pg piperazine, 8a, is added to the reaction solution. This reactionis allowed to continue at room temperature for about 1 to 24 hours. Uponcompletion of the reaction, the compound 9 is recovered by conventionalmethods including neutralization, evaporation, extraction,precipitation, chromatography, filtration, and the like, or,alternatively, is used in the next reaction without purification and/orisolation.

Conventional removal of the protecting group provides for the freepiperazine derivative, 10. Removal is accomplished in accordance withthe blocking group employed. For example, a trifluoromethylcarbonylprotecting group is readily removed via an aqueous solution of potassiumcarbonate. Further, suitable protecting groups for various functionalgroups as well as suitable conditions for protecting and deprotectingparticular functional groups are well known in the art. See, forexample, T. W. Greene and G. M. Wuts, Protecting Groups in OrganicChemistry, Second Edition, Wiley, N.Y., 1991, and references citedtherein.

The free piperazine derivative, 10, is then converted to thecorresponding carbamyl chloride, 11, by reaction in a biphasic reactionmixture of phosgene in toluene (Fluka), dichloromethane and aqueousbicarbonate solution. Subsequent reaction of the carbamyl chloride, 11,with a mono-capped PEG compound such as commercially availableCH₃(OCH₂CH₂)_(p)OH provides for PEG derivative 12. The reaction isconducted in a suitable solvent such as methylene chloride, chloroform,etc. typically in the presence of a catalytic amount of DMAP and a baseto scavenge the acid generated during reaction. The reaction iscontinued until substantially complete which typically occurs within 4to 24 hours.

When R^(a) is alkyl, subsequent hydrolysis of the ester derivativeprovides for the free carboxyl group or a salt thereof.

A specific example of this reaction scheme up to formation of thepiperazine derivative 10 is illustrated in Scheme 5 below:

Specifically, commercially available 3-pyridinesulfonic acid, 21, isconverted under conventional conditions to the corresponding sulfonylchloride, 22, by contact with POCl₃/PCl₅ using conditions well known inthe art. Coupling of sulfonyl chloride, 22, with commercially availableS-5,5-dimethylthiazolidine-4-carboxylic acid, 23, is accomplished underconventional conditions preferably in the presence of a phosphate buffer(pH 7.4) using an excess of sulfonyl chloride. The reaction ispreferably conducted at a temperature of from about −10 to 20° C. untilthe reaction is substantially complete, which typically occurs within0.5 to 5 hours. The resulting product, 24, can be recovered byconventional methods, such as chromatography, filtration, evaporation,crystallization, and the like or, alternatively, used in the next stepwithout purification and/or isolation.

The N-pyridyl sulfonyl-5,5-dimethylthiazolidine-4-carboxylic acidcompound, 23 is next coupled to t-butyl tyrosine using conventionalamino acid coupling conditions. Specifically, this coupling reaction isconducted using well known coupling reagents such as1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC),1-hydroxy-benzotriazole (HOBt) and N-methylmorpholine to facilitate thecoupling reaction.

This coupling reaction is typically conducted by contacting theN-sulfonylamino acid, 23, with about 1 to about 2 equivalents of thecoupling reagent and at least one equivalent, preferably about I toabout 1.2 equivalents, of tyrosine t-butyl ester in an inert diluent,such as dichloromethane, chloroform, acetonitrile, tetrahydrofuran,N,N-dimethylformamide and the like. Generally, this reaction isconducted at a temperature ranging from about 0° C. to about 22° C. forabout 12 to about 24 hours. Upon completion of the reaction, thecompound 24 is recovered by conventional methods includingneutralization, evaporation, extraction, precipitation, chromatography,filtration, and the like or, alternatively, is employed in the next stepwithout purification and/or isolation.

Separately, mono-N-Boc-piperazine, 25, is converted to the correspondingcarbamyl chloride, 26, by reaction with phosgene in the manner describedabove. Upon completion of the reaction, the compound 26 is recovered byconventional methods including neutralization, evaporation, extraction,precipitation, chromatography, filtration, and the like or,alternatively, is employed in the next step without purification and/orisolation.

Coupling of compound 24 with compound 26 to provide for compound 27proceeds under conventional conditions in an inert diluent such asdichloromethane, with a catalytic amount of DMAP and preferably in thepresence of a base to scavenge the acid generate. The reaction is run ata temperature of about −20 to about 22° C. for about 2 to about 24hours. Upon completion of the reaction, compound 27 is recovered byconventional methods including neutralization, evaporation, extraction,precipitation, chromatography, filtration, and the like or,alternatively, is employed in the next step without purification and/orisolation.

Removal of both the amino Boc protecting group and the t-butyl esterproceeds in the presence of trifluoroacetic acid to provide for compound28 which can be recovered by conventional methods includingneutralization, evaporation, extraction, precipitation, chromatography,filtration, and the like.

Scheme 6 below illustrates the preparation of a piperazine compoundorthogonally protected on one of the amine groups relative to thecarboxyl protecting group found on the phenylalanine compound such thatafter coupling, the piperazine protecting group can be removeddifferentially from that of the carboxyl protecting group. Suchorthogonal protection is necessary if subsequent reactions on theresulting compound require a carboxyl protecting group to avoidundesired side reactions.

Specifically, in Scheme 6, compound 24 is prepared in the mannerdescribed above. N-t-Boc-piperazine, 25, is conventionally converted toN-t-Boc-N′-trifluoromethyl-carbonylpiperazine, 29, by contact with anexcess of trifluoroacetic an hydride in the presence of a suitable aminesuch as triethylamine to scavenge the acid generated during reaction ina suitable solvent such as dichloromethane. Generally, this reaction isconducted at a temperature ranging from about −20° C. to about 22° C.for about 1 to about 24 hours. Upon completion of the reaction, compound29 can be recovered by conventional methods including neutralization,evaporation, extraction, precipitation, chromatography, filtration, andthe like or, alternatively and preferably, is employed in the next stepwithout purification and/or isolation.

In turn, removal of the t-Boc protecting group on theN-t-Boc-N′-trifluoromethylcarbonylpiperazine, 29, proceeds underconventional conditions using gaseous HCl bubbled through an inertsolvent such as methylene chloride, EtOAc, EtO₂, and the like underambient conditions to provide for the hydrochloride salt ofN′-trifluoromethylcarbonylpiperazine, 30. Generally, this reaction isconducted at a temperature ranging from about −20° C. to about 22° C.for about 0.5 to about 4 hours. Upon completion of the reaction,compound 30 can be recovered by conventional methods includingneutralization, evaporation, extraction, precipitation, chromatography,filtration, and the like or, alternatively and preferably, is employedin the next step without purification and/or isolation.

Conversion of N′-trifluoromethylcarbonylpiperazine, 30, to theN-carbamyl chloride derivative, 31, conventionally proceeds by contactwith phosgene in the manner described above. Upon completion of thereaction, compound 31 can be recovered by conventional methods includingneutralization, evaporation, extraction, precipitation, chromatography,filtration, and the like or, alternatively and preferably is employed inthe next step without purification and/or isolation.

Compounds 31 and 24 are coupled under conditions similar to thosedescribed above to provide for compound 32 which is orthogonallyprotected at the amino moiety of the piperazine group as well as thecarboxyl moiety of the phenylalanine group. Selective removal of thetrifluoromethylcarbonyl amino protecting group proceeds underconventional conditions using an aqueous solution of potassium carbonateto provide for compound 33.

Scheme 7 below illustrates a first route for derivatization of compound28 to provide for PEG substitution. In this scheme, the amino moiety ofthe piperazine group is employed as a complementary functional group tothe activated carboxyl group of the lysine derivative to form a covalentamide bond thereby introducing two PEG moieties into the compoundthrough a linker of the formula

which linker comprises 8 carbon atoms and 5 heteroatoms.

Specifically, in Scheme 7, conjugation of an excess of compound 28 (1.1to 10 eq) with commercially available N-hydroxysuccinimidyl ester of adi-PEG substituted lysine derivative, in the presence of phosphatebuffered aqueous solution provides for compound 29 which is recovered bydialysis. The commercially available N-hydroxy-succinimidyl ester of adi-PEG substituted lysine derivative has a weight average molecularweight of about 40,000 which means that each PEG moiety has a numberaverage molecular weight of about 20,000. The reaction is run at atemperature of about 0 to about 22° C.

Scheme 8 illustrates a second route for derivatization to provide forPEG substitution. In this scheme, the amino moiety of the piperazinegroup is employed as a complementary functional group to an in situformed activated carboxyl group of a commercially available carboxyl-PEGcompound which under conventional reactive conditions forms a covalentamide bond thereby introducing a single PEG moiety into the compound. Inthis embodiment, the carboxyl-PEG compound is represented by the formulaHOOC(CH₂)_(v)(OCH₂CH₂)_(p)OCH₃ where p and v are as defined above andthe resulting linker to the PEG group is represented by —C(O)(CH₂)_(v).Carboxylated PEG compounds can be made by oxidation of the hydroxyterminated PEG compounds using conventional methods and reagents.

Specifically, in Scheme 8, an excess (1.1 to 10 equiv) of compound 33,prepared as in Scheme 7, is added to at least an equivalent of acommercially available carboxyl-PEG compound which is convertd in situto an activated ester (not shown) by contact with at least an equivalentand preferably an excess of HATU[O-(7-azabenzotriazol-1-yl)-1,1,3,3,-tetramethyluroniumhexafluorophosphate] in the presence of a suitable amine such astriethylamine. Coupling of the carboxyl-PEG compound to compound 33preferably proceeds at a temperature of from about 0 to about 22° C. forabout 2 to about 24 hours. Upon completion of the reaction, the compound34 is recovered by conventional methods including neutralization,evaporation, extraction, precipitation, chromatography, filtration, andthe like or, alternatively, is employed in the next step withoutpurification and/or isolation.

Conventional removal of the t-butyl carboxyl protecting group with anexcess of formic acid provides for a mono-PEG compound of formula XXIIof this invention.

Scheme 9 illustrates a third route for derivatization to provide for PEGsubstitution. In this scheme, the amino moiety of the piperazine groupis employed as a complementary functional group to an in situ formedchloroformate of a commercially available mono-hydroxy-PEG compoundwhich under conventional reactive conditions forms a covalent carbamatebond thereby introducing a single PEG moiety into the compound. In thisembodiment, the mono-hydroxy-PEG compound is represented by the formulaHOCH₂CH₂(OCH₂CH₂)_(p)OCH₃ where p is as defined above and the resultinglinker is represented by —C(O)—.

Specifically, in Scheme 9, the hydroxyl group of a commerciallyavailable mono-hydroxy PEG, 36, is converted to the correspondingchloroformate, 37, by reaction with phosgene in toluene (Fluka), indichloromethane. The product is isolated by evaporation and is employedin the next step without further purification.

A slight excess (1.1 to 10 eq) of chloroformate 37 is contacted withcompound 33, prepared as above, in the presence of a suitable base suchas triethylamine to scavenge the acid generated. Coupling of thechloroformate-PEG compound to compound 33 preferably proceeds at atemperature of from about 0 to about 22° C. for about 2 to about 4hours. Upon completion of the reaction, the compound 38 is recovered byconventional methods including neutralization, evaporation, extraction,precipitation, chromatography, filtration, and the like or,alternatively, is employed in the next step without purification and/orisolation.

Conventional removal of the t-butyl carboxyl protecting group with anexcess of formic acid provides for a mono-PEG compound, 39, of formulaXXII of this invention.

Scheme 10 illustrates the synthesis of two intermediates useful forsubsequent PEG substitution. In this scheme, the amino moiety of thepiperazine group is employed as a complementary functional group whichis derivatized for subsequent PEG substitution.

Specifically, in Scheme 10, conversion of amino moiety of the piperazinegroup to the corresponding N-carbamyl chloride derivative, 40, proceedsby contact with an excess of phosgene in the presence of a suitable basesuch as sodium bicarbonate to scavenge the acid generated duringreaction. Upon completion of the reaction, compound 40 can be recoveredby conventional methods including neutralization, evaporation,extraction, precipitation, chromatography, filtration, and the like or,alternatively and preferably, is employed in the next step (illustratedin Scheme 11) without purification and/or isolation.

Alternatively, the amino moiety of the piperazine group of compound 33can be converted to the corresponding amide, compound 41, by reactionwith at least an equivalent and preferably an excess of 4-nitrobenzoylchloride in the presence of a base such as pyridine (which can also actas a solvent) to scavenge the acid generated during reaction. Thereaction preferably proceeds at a temperature of from about 0 to about22° C. for about 1 to about 24 hours. Upon completion of the reaction,compound 41 is recovered by conventional methods includingneutralization, evaporation, extraction, precipitation, chromatography,filtration, and the like or, alternatively, is employed in the next stepwithout purification and/or isolation.

Subsequent reduction of the para-nitro substituent of the phenyl groupprovides for the amine substituent in compound 42. Reduction isconventionally conducted using palladium/carbon under a hydrogenatmosphere.typically at elevated pressures in a suitable diluent such asmethanol. The reaction-proceeds until-substantial completion whichtypically occurs within about 24 to about 72 hours. During the reaction,additional catalyst is added as required to affect reaction completion.Upon completion of the reaction, the compound 42 is recovered byconventional methods including neutralization, evaporation, extraction,precipitation, chromatography, filtration, and the like or,alternatively, is employed in the next step without purification and/orisolation.

Conversion of the para-amino substituent of the phenyl group of compound42 to the corresponding isocyanate, 43, occurs by reaction with anexcess of phosgene in the presence of a suitable base such as sodiumbicarbonate which scavenges the acid generated. The reaction proceedsuntil substantial completion which typically occurs within about 0.5 toabout 5 hours at about 0° C. to about 22° C. Upon completion of thereaction, the compound 43 is recovered by conventional methods includingneutralization, evaporation, extraction, precipitation, chromatography,filtration, and the like or, alternatively, is employed in the next stepwithout purification and/or isolation.

Scheme 11 illustrates a fourth route for derivatization to provide forPEG substitution. In this scheme, the carbamyl chloride moiety of thepiperazine group of compound 40 is employed as a complementaryfunctional group to form a carbamate or urea bond with a commerciallyavailable mono-hydroxy- or mono-amino-PEG compound which underconventional reactive conditions. In this embodiment, the PEG compoundis represented by the formula HQCH₂CH₂(OCH₂CH₂)_(p)OCH₃ where p and Qare as defined above and the resulting linker is represented by —C(O)—.

Specifically, in Scheme 11, an excess (1.1 to 10 eq) of carbamylchloride, 40, is contacted in an inert solvent such as dichloromethanewith a suitable mono-hydroxy- or mono-amino-PEG compound preferably inthe presence of a suitable base such as triethylamine and/or catalyticamounts of 4-N,N-dimethylaminopyridine (DMAP). The reaction proceedsuntil substantial completion which typically occurs within about 4 toabout 48 hours. Upon completion of the reaction, the compound 44 isrecovered by conventional methods including neutralization, evaporation,extraction, precipitation, chromatography, filtration, and the like or,alternatively, is employed in the next step without purification and/orisolation.

When Q is a hydroxyl group, the resulting product contains a carbamatefunctionality covalently linking the PEG group to the VLA-4 antagonistthrough a linker represented by —C(O)—. When Q is an amino group, theresulting product contains a urea functionality covalently linking thePEG group to the VLA-4 antagonist through a linker represented by—C(O)—.

Conventional removal of the t-butyl carboxyl protecting group with anexcess of formic acid provides for a mono-PEG compound, 45, of formulaXXIIa of this invention.

Scheme 12 illustrates a fifth route for derivatization to provide forPEG substitution. In this scheme, the isocyanate moiety of the phenylgroup of compound 43 is employed as a complementary functional group toform a carbamate or urea bond with a commercially availablemono-hydroxy- or mono-amino-PEG compound which under conventionalreactive conditions. In this embodiment, the PEG compound is representedby the formula HQCH₂CH₂(OCH₂CH₂)_(p)OCH₃ where p and Q are as definedabove and the resulting linker is represented by:

where the linker comprises 8 carbon atoms and 3 heteroatoms.

Specifically, in Scheme 12, an excess (1.1 to 10 eq) isocyanate, 43, iscontacted with a suitable mono-hydroxy- or mono-amino-PEG compound in asuitable inert diluent such as dichloromethane or toluene. The reactionis preferably maintained at a temperature of from about 0° to about 105°C. until substantial completion which typically occurs within about 1 toabout 24 hours. Upon completion of the reaction, compound 46 isrecovered by conventional methods including neutralization, evaporation,extraction, precipitation, chromatography, filtration, and the like or,alternatively, is employed in the next step without purification and/orisolation.

When Q is a hydroxyl group, the resulting product contains a carbamatefunctionality covalently linking the PEG group to the VLA-4 antagonistthrough a —C(O)— linking group. When Q is an amino group, the resultingproduct contains a urea functionality covalently linking the PEG groupto the VLA-4 antagonist through a —C(O)— linking group.

Conventional removal of the t-butyl carboxyl protecting group with anexcess of formic acid provides for a mono-PEG compound, 47, of formulaXXII of this invention.

In the Schemes above, amine moieties located on other portions of themolecule can be employed in the manner described above to covalentlylink a PEG group to the molecule. For example, amines located on Ar¹, onthe heterocyclic amino acid or on Ar² can be similarly derivatized toprovide for PEG substitution. The amine moieties can be included inthese substituents during synthesis and appropriately protected asnecessary. Alternatively, amine precursors can be employed. For example,as shown in Scheme 10, reduction of a nitro group provides for thecorresponding amine. Similarly, reduction of a cyano group provides fora H₂NCH₂— group. Nitro and cyano substituted Ar¹ groups are provided inU.S. Pat. No. 6,489,300 as is an amino substituted Ar¹ group.

Further, the amino substitution can be incorporated into theheterocyclic amino acid functionality and then derivatized to include aPEG moiety found in formula XXII as R. For example, the heterocyclicamino acid functionality can be 2-carboxylpiperazine depicted in U.S.Pat. No. 6,489,300. Alternatively, commercially available 3- or4-hydroxyproline can be oxidized to the corresponding ketone and thenreductively aminated with ammonia in the presence of sodiumcyanoborohydride to form the corresponding amine moiety. Still further,4-cyanoproline can be reduced to provide for a substituted alkyl groupof the formula CH₂NH₂ which can be derivatized through the amine.

Still further, the amine moiety can be incorporated into the Ar²functionality. Preferably, the amine moiety is present as an amineprecursor such as a nitro or cyano group bound to Ar².

In the schemes above, the reactions of the amine with a complementaryfunctional group can be reversed such that the carboxyl or hydroxylgroup is on the VLA-4 antagonist of formula XXIIa (without any PEGsubstituents) and the amine group could be part of the PEG moiety. Insuch cases, the amine group, preferably terminating the PEG moiety, canbe converted to an isocyanate, using phosgene and Et₃N, and reacted withthe hydroxyl group to form a carbamate as illustrated in Scheme 13below:

Specifically, compound 48 described in U.S. Pat. No. 6,489,300 iscontacted with at least an equivalent and preferably an excess of 49 inthe manner described above to provide for the corresponding carbamate,50. Deprotection, as described above, then provides for compound 51.

Alternatively, in Scheme 13, the hydroxyl functionality can be reactedwith phosgene 10 to provide for the chlorocarbonyloxy derivative whichreacts with an amine group of a monoamine compound to provide for thecarbamate.

Carboxyl functionality, for example on the Ar¹ moiety, can be convertedto the corresponding amide by reaction with a mono-amino-PEG compound inthe manner described above in Scheme 8.

Specifically, in Scheme 14, known compound 52, described in U.S. Pat.No. 6,489,300, is t-butyl protected under convention conditions toprovide the cyano compound 53, which is hydrogenated under conventionalconditions to provide the aminomethyl compound 54. The aminomethyl groupis reacted with Et₃N and a PEG chloroformate, as illustrated previouslyin Scheme 9, to provide the carbamate-linked conjugate t-butyl ester 55.Treatment of the t-butyl ester with HCO₂H provides the conjugatecarboxylic acid 56.

Suitable PEG compounds are commercially available or can be prepared byart recognized procedures. For example, mono-capped linear PEGs with oneterminal amine are available in varying molecular weights (e.g., 2kilodaltons (kDa), 5 kDa, 10 kDa and 20 kDa from Nektar, San Carlos,Calif.). Preferred mono-capped PEGs having one terminal amine group canbe represented by the formula H₂NCH₂CH₂(OCH₂CH₂)_(p)OCH₃.

Mono-capped linear PEGs with one terminal alcohol are available invarying molecular weights (e.g., 2 kilodaltons (kDa), 5 kDa, 10 kDa and20 kDa from Nektar, San Carlos, Calif.). Preferred mono-capped linearPEGs having one terminal alcohol can be represented by the formulaHOCH₂CH₂(OCH₂CH₂)_(p)OCH₃.

Diamino-capped linear PEGs having an amino group at both termini arecommercially available and are sometimes referred to as “Jeffamines”(tradename of Huntsman). Preferred diamino-capped linear PEGs having anamino group at both termini can be represented by the formula:H₂NCH₂CH₂(OCH₂CH₂)_(p)NH₂.

Scheme 15 below illustrates an alternative synthesis of3-aminopyrrolidinyl derivatives useful as starting materials in thisinvention for subsequent PEG substitution at the amino group.

Using conventional methods, commercially available cis-4-hydroxyL-proline, 57, is treated with methanolic hydrogen chloride for severalhours at reflux, followed by evaporation, and the so generated methylester hydrochloride is treated with excess tosyl chloride in pyridinefor two days at room temperature, giving the product, 58. Compound 58 isisolated by neutralizing the pyridine using weak aqueous acid andextracting the product with an organic solvent such as EtOAc. Theproduct 58 may be purified by crystallization, flash chromatography, ormore preferably be used in subsequent steps without purification.

Reaction of 58 with a saturated solution of excess sodium azide in DMFat room temperature for 15 days affords compound 59. Compound 59 isisolated by dilution of the reaction mixture with water, followed byextraction with an organic solvent such as EtOAc. The product 59 may bepurified by crystallization, flash chromatography, or more preferably beused in subsequent steps without purification.

Compound 59 is treated with sodium hydroxide, in a mixture of water andmethanol, thus hydrolyzing the methyl ester and generating a carboxylicacid, which is isolated by acidification and extraction with an organicsolvent such as EtOAc. The carboxylic acid is treated with L-tyrosinet-butyl ester [H-Tyr(H)-OtBu], EDAC, HOBt, and Et3N in DMF, generating adipeptide, which is isolated by dilution with water and extraction withan organic solvent such as EtOAc. The dipeptide is treated withClCONMe2, Et3N, and DMAP in DCM at reflux for 24 hours, generating thecarbamate, 60, which is isolated by dilution with EtOAc, sequentialwashing with weak aqueous acid and base, and then evaporation. Compound60 is rigorously purified by flash chromatography.

Finally, compound 61 is prepared by shaking of a solution of 60 inmethanol, with a Pd/C catalyst under an atmosphere of hydrogen. Theproduct, 61, is isolated by removal of the catalyst by filtration andevaporation.

Still further, the synthesis of varying mono-capped mono-hydroxy PEGsare described in detail by Campbell, U.S. Pat. No. 4,604,103 which isincorporated herein by reference in its entirety. If a mono-cappedmono-amino PEG is preferred, the mono-capped mono-hydroxy PEGs canreadily be converted to the corresponding chloride by conventionalmethods and subsequently converted to an amine by contact with an excessof ammonia.

The PEGs of this invention comprise, for example, the following:

-   -   HO(alkylene-O)_(p)H dihydroxy-PEG    -   HO(alkylene-O)_(p)R^(b) mono-capped mono-hydroxy PEG    -   H₂N(alkylene-O)_(p)R^(b) mono-capped mono-amino PEG    -   H₂N(alkylene-O)_(p)CH₂CH₂NH₂ Jeffamines        where p and alkylene are as defined herein and R^(b) is        preferably selected from the group consisting of alkyl,        substituted alkyl, aryl and substituted aryl.

The PEG derivatives described herein can be used in the pharmaceuticalsformulations described above. Preferably, the formulations areadministered orally or parenterally to a subject in need thereof.

Antibodies & Immunoglobulins

In one specific embodiment, the agents of the invention areimmunoglobulins that selectively bind to an alpha-4 integrin or a dimercomprising alpha-4 integrin, such as alpha-4 beta-1 or alpha-4 beta-7.The immunoglobulins are preferably antibodies or fragments thereof thatbind to an alpha-4 integrin or dimer thereof. Also contemplated hereinare immunoglobulin molecules that bind to VCAM- 1 in a manner such thatthey inhibit VCAM- 1 interaction with VLA-4. By antibodies is meant toinclude complete immunoglobulins such as IgG1 or IgM, or inhibitorsderived from antibodies, such as Antegren™. Preferably, theimmunoglobulins recognize epitopes on VLA-4 and by recognizing andbinding to these epitopes; the immunoglobulines inhibit VLA-4 frominteracting with VCAM-1.

When the agent of the invention is an antibody, a monoclonal antibody isthe preferred antibody. In contrast to polyclonal antibody preparations,which typically include different antibodies directed against differentepitopes, each monoclonal antibody is directed against a single epitopeon the antigen. A second advantage of monoclonal antibodies is that theyare synthesized by means that are uncontaminated by otherimmunoglobulins, e.g., by phage display or isolation from a hybridoma.Although the present invention intends to encompass both polyclonal andmonoclonal antibodies as agents of the invention, monoclonal antibodiesare preferred as they are highly specific, and the invention is thusdiscussed primarily in terms of monoclonal antibodies.

“Native antibodies and immunoglobulins” are usually heterotetramericglycoproteins of about 150,000 Daltons, composed of two identical light(L) chains and two identical heavy (H) chains. Each light chain islinked to a heavy chain by one covalent disulfide bond, while the numberof disulfide linkages varies between the heavy chains of differentimmunoglobulin isotypes. Each heavy and light chain also has regularlyspaced. intrachain disulfide bridges. Each heavy chain has at one end avariable domain (V_(H)) followed by a number of constant domains. Eachlight chain has a variable domain at one and (V_(L)) and a constantdomain at its other end; the constant domain of the light chain isaligned with the first constant domain of the heavy chain, and the lightchain variable domain is aligned with the variable domain of the heavychain. Particular amino acid residues are believed to form an interfacebetween the light and heavy chain variable domains (Clothia et al.,1985, J. Mol. Biol., 186: 651-63; Novotny et al., 1985, Proc. Natl.Acad. Sci. USA, 82: 4592-6).

In addition, other antibodies can be identified using techniquesavailable in the art. For example, monoclonal antibodies of the presentinvention can be produced using phage display technology. Antibodyfragments, which selectively bind to an alpha-4 integrin or a dimercomprising an alpha-4 integrin, are then isolated. Exemplary preferredmethods for producing such antibodies via phage display are disclosed inU.S. Pat. Nos. 6,225,447; 6,180,336; 6,172,197; 6,140,471; 5,969,108;5,885,793; 5,872,215; 5,871,907; 5,858,657; 5,837,242; 5,733,743 and5,565,332.

A “variant” antibody, refers herein to an immunoglobulin molecule thatdiffers in amino acid sequence from a “parent” antibody amino acidsequence by virtue of addition, deletion and/or substitution of one ormore amino acid residue(s) in the parent antibody sequence. The parentantibody or immunoglobulin can be a polyclonal antibody, monoclonalantibody, humanized antibody, Primatized® antibody or any antibodyfragment. In the preferred embodiment, the variant comprises one or moreamino acid substitution(s) in one or more hypervariable region(s) of theparent antibody. For example, the variant may comprise at least one,e.g., from about one to about ten, and preferably from about two toabout five, substitutions in one or more hypervariable regions of theparent antibody. Ordinarily, the variant will have an amino acidsequence having at least 75% amino acid sequence identity with theparent antibody heavy or light chain variable domain sequences, morepreferably at least 80%, more preferably at least 85%, more preferablyat least 90%, and most preferably at least 95%. Identity or homologywith respect to this sequence is defined herein as the percentage ofamino acid residues in the candidate sequence that are identical withthe parent antibody residues, after aligning the sequences andintroducing gaps, if necessary, to achieve the maximum percent sequenceidentity. No N-terninal, C-terminal, or internal extensions, deletions,or insertions into the antibody sequence shall be construed as affectingsequence identity or homology. The variant retains the ability to bindthe receptor and preferably has properties that are superior to those ofthe parent antibody. For example, the variant may have a strongerbinding affinity, enhanced ability to activate the receptor, etc. Toanalyze such properties, one should compare a Fab form of the variant toa Fab form of the parent antibody or a full-length form of the variantto a full-length form of the parent antibody. The variant antibody ofparticular interest herein is one which displays at least about 10 fold,preferably at least about 20 fold, and most preferably at least about 50fold, enhancement in biological activity when compared to the parentantibody. The “parent” antibody herein is one that is encoded by anamino acid sequence used for the preparation of the variant. Preferably,the parent antibody has a human framework region and has human antibodyconstant region(s). For example, the parent antibody may be a humanizedor human antibody. An “isolated” antibody is one that has beenidentified and separated and/or recovered from a component of itsnatural environment. Contaminant components of its natural environmentare materials that would interfere with diagnostic or therapeutic usesfor the antibody, and may include enzymes, hormones, and otherproteinaceous or non-proteinaceous solutes. In preferred embodiments,the antibody will be purified (1) to greater than 95% by weight ofantibody as determined by the Lowry method, and most preferably morethan 99% by weight, (2) to a degree sufficient to obtain at least 15residues of N-terminal or internal amino acid sequence by use of aspinning cup sequenator, or (3) to homogeneity by SDS-PAGE underreducing or non-reducing conditions using Coomassie blue or, preferably,silver stain. Isolated antibody includes the antibody in situ withinrecombinant cells since at least one component of the antibody's naturalenvironment will not be present. Ordinarily, however, isolatedantibodies will be prepared by at least one purification step.

Monoclonal Antibodies. Monoclonal antibodies can also be produced usingthe conventional hybridoma methods. These methods have been widelyapplied to produce hybrid cell lines that secrete high levels ofmonoclonal antibodies against many specific antigens, and can also beused to produce monoclonal antibodies of the present invention. Forexample, mice (e.g., Balb/c mice) can be immunized with an antigenicalpha-4 epitope by intraperitoneal injection. After sufficient time haspassed to allow for an immune response, the mice are sacrificed and thespleen cells obtained and fused with myeloma cells, using techniqueswell known in the art. The resulting fused cells, hybridomas, are thengrown in a selective medium, and the surviving cells grown in suchmedium using limiting dilution conditions. After cloning and recloning,hybridomas can be isolated that secrete antibodies (for example, of theIgG or IgM class or IgGI subclass) that selectively bind to the target,alpha-4 or a dimer comprising an alpha-4 integrin. To produce agentsspecific for human use, the isolated monoclonal can then be used toproduce chimeric and humanized antibodies. Antibodies can also beprepared that are anti-peptide antibodies. Such anti-peptide antibodieswould be prepared against peptides of alpha-4 integrin.

Chimeric, Primatized® and humanized antibodies can be produced fromnon-human antibodies, and can have the same or similar binding affinityas the antibody from which they are produced. Techniques developed forthe production of chimeric antibodies (Morrison et al., 1984 Proc. Natl.Acad. Sci. 81: 6851; Neuberger et al., 1984 Nature 312: 604; Takeda etal., 1985 Nature 314: 452) by splicing the genes from a mouse antibodymolecule of appropriate antigen specificity together with genes from,for example, a human antibody molecule of appropriate biologicalactivity can be used; such antibodies are within the scope of thisinvention. For example, a nucleic acid encoding a variable (V) region ofa mouse monoclonal antibody can be joined to a nucleic acid encoding ahuman constant (C) region, e.g., IgG1 or IgG4. The resulting antibody isthus a species hybrid, generally with the antigen binding domain fromthe non-human antibody and the C or effector domain from a humanantibody.

Humanized antibodies are antibodies with variable regions that areprimarily from a human antibody (the acceptor antibody), but which havecomplementarity detennining regions substantially from a non-humanantibody (the donor antibody). See, e.g., Queen et al., 1989 Proc.NatlAcad. Sci. USA 86: 10029-33; WO 90/07861; and U.S. Pat. Nos.6,054,297; 5,693,761; 5,585,089; 5,530,101 and 5,224,539. The constantregion or regions of these antibodies are generally also from a humanantibody. The human variable domains are typically chosen from humanantibodies having sequences displaying a high homology with the desirednon-human variable region binding domains. The heavy and light chainvariable residues can be derived from the same antibody, or a differenthuman antibody. In addition, the sequences can be chosen as a consensusof several human antibodies, such as described in WO 92/22653.

Specific amino acids within the human variable region are selected forsubstitution based on the predicted conformation and antigen bindingproperties. This can be determined using techniques such as computermodeling, prediction of the behavior and binding properties of aminoacids at certain locations within the variable region, and observationof effects of substitution. For example, when an amino acid differsbetween a non-human variable region and a human variable region, thehuman variable region can be altered to reflect the amino acidcomposition of the non-human variable region.

In a specific embodiment, the antibodies used in the chronic dosageregime of the present invention are humanized antibodies as disclosed inU.S. Pat. No. 5,840,299, which is incorporated herein by reference.

In another embodiment, transgenic mice containing human antibody genescan be immunized with an antigenic alpha-4 structure and hybridomatechnology can be used to generate human antibodies that selectivelybind to alpha-4.

Chimeric, human and/or humanized antibodies can be produced byrecombinant expression, e.g., expression in human hybridomas (Cole etal., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77(1985)), in myeloma cells or in Chinese Hamster Ovary (CHO) cells.Alternatively, antibody-coding sequences can be incorporated intovectors suitable for introducing into the genome of animal therebyproducing a transgenic animal. One example would be to produce suchantibodies in the milk of a transgenic animal such as a bovine. See,e.g., U.S. Pat Nos. 5,849,992 and 5,304,489. Suitable transgenes includetrangenes having a promoter and/or enhancer from a mammary glandspecific gene, for example casein or β-lactoglobulin.

Natalizumab And Related Humanized Antibodies

The invention provides for a method of using humanized immunoglobulinsthat specifically bind to a VLA-4 ligand either alone or in combinationto diagnose and/or treat rheumatoid arthritis. One preferred antibodyfor use in such methods of treatment and in medicaments includes thatdescribed in U.S. Pat. No. 5,840,299 assigned to Elan Pharmaceuticals,which is herein incorporated in its entirety. Another aspectcontemplates the use of fragments of these antibodies as assessed invivo.

The humanized antibodies comprise a humanized light chain and ahumanized heavy chain. In one aspect, the humanized light chain cancomprise three complementarity determining regions (i.e., CDR1, CDR2 andCDR3) having amino acid sequences from the corresponding complementaritydetermining regions of a mouse 21-6 immunoglobulin light chain, and avariable region framework from a human kappa light chain variable regionframework sequence except in at least one position selected from a firstgroup consisting of positions L45, L49, L58 and L69, wherein the aminoacid position is occupied by the same amino acid present in theequivalent position of the mouse 21.6 immunoglobulin light chainvariable region framework.

The humanized heavy chain comprises three complementarity determiningregions (i.e., CDR1, CDR2 and CDR3) having amino acid sequences from thecorresponding complementarity determining regions of a mouse 21-6immunoglobulin heavy chain, and a variable region framework from a humanheavy chain variable region framework sequence except in at least oneposition selected from a group consisting of H27, H28, H29, H30, H44,H71, wherein the amino acid position is occupied by the same amino acidpresent in the equivalent position of the mouse 21-6 immunoglobulinheavy chain variable region framework. The immunoglobulins specificallybind to VLA-4 with an affinity having a lower limit of about 10⁷ M⁻¹ andan upper limit of about five times the affinity of the mouse 21-6immunoglobulin.

Usually, the humanized light and heavy chain variable region frameworksare from RE1 and 21/28′CL variable region framework sequencesrespectively. When the humanized light chain variable region frameworkis from RE1, at least two framework amino acids are replaced. One aminoacid is from the first group of positions described supra. The otheramino acids are from a third group consisting of positions L104, L105and-L107. This position is occupied by the same amino acid present inthe equivalent position of a kappa light chain from a humanimmunoglobulin other than RE1.

Some humanized immunoglobulins have a mature light chain variable regionsequence designated La or Lb, or a mature heavy chain variable regionsequence designated Ha, Hb or Hc (FIG. 13). Preferred humanizedimmunoglobulins include those having a La light chain and an Ha, Hb orHe heavy chain.

The humanized immunoglobulins have variable framework regionssubstantially from a human immunoglobulin (termed an acceptorimmunoglobulin) and complementarity determining regions substantiallyfrom a mouse immunoglobulin termed mu MAb 21.6 (referred to as the donorimmunoglobulin). The constant region(s), if present, are alsosubstantially from a human immunoglobulin. The humanized antibodiesexhibit a specific binding affinity for VLA-4 of at least 10⁷, 10⁸, 10⁹,or 10¹⁰ M⁻¹. Usually the upper limit of binding affinity of thehumanized antibodies for VLA-4 is within a factor of three or five ofthat of mu MAb 21.6 (about 10⁹ M⁻¹). Often the lower limit of bindingaffinity is also within a factor of three or five of that of mu MAb21.6.

Humanized antibodies can be produced as exemplified, for example, withthe mouse MAb 21.6 monoclonal antibody. The starting material forproduction of humanized antibodies is mu MAb 21.6. The isolation andproperties of this antibody are described in U.S. Pat. No. 6,033,655(assigned to Elan Pharmaceuticals, Inc.), which is herein incorporatedby reference in its entirety. Briefly, mu MAb 21.6 is specific for theα₄ subunit of VLA-4 and has been shown to inhibit human lymphocytebinding to tissue cultures of rat brain cells stimulated with tumornecrosis factor. From N-terminal to C-terminal, both light and heavychains comprise the domains FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. Theassignment of amino acids to each domain is in accordance with thenumbering convention of Kabat.

The next step involved selecting human antibodies to supply frameworkresidues. The substitution of mouse CDRs into a human variable domainframework is most likely to result in retention of their correct spatialorientation if the human variable domain framework adopts the same orsimilar conformation to the mouse variable framework from which the CDRsoriginated. This is achieved by obtaining the human variable domainsfrom human antibodies whose framework sequences exhibit a high degree ofsequence identity with the murine variable framework domains from whichthe CDRs were derived. The heavy and light chain variable frameworkregions can be derived from the same or different human antibodysequences. The human antibody sequences can be the sequences ofnaturally occurring human antibodies or can be consensus sequences ofseveral human antibodies. See Kettleborough et al., Protein Engineering4: 773 (1991); Kolbinger et al., Protein Engineering 6: 971 (1993).

Suitable human antibody sequences are identified by computer comparisonsof the amino acid sequences of the mouse variable regions with thesequences of known human antibodies. The comparison is performedseparately for heavy and light chains but the principles are similar foreach. This comparison reveals that the mu 21.6 light chain showsgreatest sequence identity to human light chains of subtype kappa 1; themu 21.6 heavy chain shows greatest sequence identity to human heavychains of subtype one, as defined by Kabat, supra. Thus, light and heavyhuman framework regions are usually derived from human antibodies ofthese subtypes, or from consensus sequences of such subtypes. Thepreferred light and heavy chain human variable regions showing greatestsequence identity to the corresponding regions from mu MAb 21.6 are fromantibodies RE1 and 21/28′CL respectively.

Computer modeling can then be used to further enhance the humanizedantibody's ability to bind to its cognate antigen. The unnaturaljuxtaposition of murine CDR regions with human variable framework regioncan result in unnatural conformational restraints, which, unlesscorrected by substitution of certain amino acid residues, lead to lossof binding affinity. The selection of amino acid residues forsubstitution is determined, in part, by computer modeling. Computerhardware and software for producing three-dimensional images ofimmunoglobulin molecules are widely available. In general, molecularmodels are produced starting from solved structures for immunoglobulinchains or domains thereof. The chains to be modeled are compared foramino acid sequence similarity with chains or domains of solved threedimensional structures, and the chains or domains showing the greatestsequence similarity is/are selected as starting points for constructionof the molecular model. For example, for the light chain of mu MAb 21.6,the starting point for modeling the framework regions, CDR1 and CDR2regions, was the human light chain RE1. For the CDR3 region, thestarting point was the CDR3 region from the light chain of a differenthuman antibody HyHEL-5. The solved starting structures are modified toallow for differences between the actual amino acids in theimmunoglobulin chains or domains being modeled, and those in thestarting structure. The modified structures are then assembled into acomposite immunoglobulin. Finally, the model is refined by energyminimization and by verifying that all atoms are within appropriatedistances from one another and that bond lengths and angles are withinchemically acceptable limits.

As noted supra, the humanized antibodies of the invention comprisevariable framework regions substantially from a human immunoglobulin andcomplementarity determining regions substantially from a mouseimmunoglobulin termed mu MAb 21.6. Having identified the complementaritydetermining regions (CDRs) of mu MAb 21.6 and appropriate human acceptorimmunoglobulins, the next step is to determine which, if any, residuesfrom these components should be substituted to optimize the propertiesof the resulting humanized antibody. In general, substitution of humanamino acid residues with murine should be minimized, becauseintroduction of murine residues increases the risk of the antibodyeliciting a HAMA response in humans. Amino acids are selected forsubstitution based on their possible influence on CDR conformationand/or binding to antigen. Investigation of such possible influences isby modeling, examination of the characteristics of the amino acids atparticular locations, or empirical observation of the effects ofsubstitution or mutagenesis of particular amino acids.

When an amino acid differs between a mu MAb 21.6 variable frameworkregion and an equivalent human variable framework region, the humanframework amino acid should usually be substituted by the equivalentmouse amino acid if it is reasonably expected that the amino acid:

-   -   (1) non-covalently binds antigen directly (e.g., amino acids at        positions L49, L69 of mu MAb 21.6),    -   (2) is adjacent to a CDR region, is part of a CDR region under        the alternative definition proposed by Chothia et al., supra, or        otherwise interacts with a CDR region (e.g., is within about 3 Å        of a CDR region) (e.g., amino acids at positions L45, L58, H27,        H28, H29, H30 and H71 of mu MAb 21.6), or    -   (3) participates in the VL-VH interface (e.g., amino acids at        position H44 of mu MAb 21.6).

Other candidates for substitution are acceptor human framework aminoacids that are unusual for a human immunoglobulin at that position(e.g., amino acids at positions L104, L105 and L107 of mu MAb 21.6).These amino acids can be substituted with amino acids from theequivalent position of more typical human immunoglobulins.Alternatively, amino acids from equivalent positions in the mouse MAb21.6 can be introduced into the human framework regions when such aminoacids are typical of human immunoglobulin at the equivalent positions.

In general, substitution of all or most of the amino acids fulfillingthe above criteria is desirable. Occasionally, however, there is someambiguity about whether a particular amino acid meets the abovecriteria, and alternative variant immunoglobulins are produced, one ofwhich has that particular substitution, the other of which does not. Thehumanized antibodies will usually contain a substitution of a humanlight chain framework residue with a corresponding mu MAb 21.6 residuein at least 1, 2 or 3, and more usually 4, of the following positions:L45, L49, L58 and L69. The humanized antibodies also usually contain asubstitution of a human heavy chain framework residue in at least 1, 2,3, 4, or 5, and sometimes 6, of the following positions: H27, H28, H29,H30, H44 and H71. Optionally, H36 may also be substituted. In preferredembodiments when the human light chain acceptor immunoglobulin is RE1,the light chain also contains substitutions in at least 1 or 2, and moreusually 3, of the following positions: L104, L105 and L107. Thesepositions are substituted with the amino acid from the equivalentposition of a human immunoglobulin having a more typical amino acidresidues. Appropriate amino acids to substitute are shown in FIGS. 13and 14.

Usually the CDR regions in humanized antibodies are substantiallyidentical, and more usually, identical to the corresponding CDR regionsin the mu MAb 21.6 antibody. Occasionally, however, it is desirable tochange one of the residues in a CDR region. For example, Example 4identifies an amino acid similarity between the mu MAb 21.6 CDR3 and theVCAM-1 ligand. This observation suggests that the binding affinity ofhumanized antibodies might be improved by redesigning the heavy chainCDR3 region to resemble VCAM-1 even more closely. Accordingly, one ormore amino acids from the CDR3 domain can be substituted with aminoacids from the VCAM-1 binding domain. Although not usually desirable, itis sometimes possible to make one or more conservative amino acidsubstitutions of CDR residues without appreciably affecting the bindingaffinity of the resulting humanized immunoglobulin.

Other than for the specific amino acid substitutions discussed above,the framework regions of humanized immunoglobulins are usuallysubstantially identical, and more usually, identical to the frameworkregions of the human antibodies from which they were derived. Of course,many of the amino acids in the framework region make little or no directcontribution to the specificity or affinity of an antibody. Thus, manyindividual conservative substitutions of framework residues can betolerated without appreciable change of the specificity or affinity ofthe resulting humanized immunoglobulin. However, in general, suchsubstitutions are undesirable.

Production of Variable Regions.

Having conceptually selected the CDR and framework components ofhumanized immunoglobulins, a variety of methods are available forproducing such immunoglobulins. Because of the degeneracy of the code, avariety of nucleic acid sequences will encode each immunoglobulin aminoacid sequence. The desired nucleic acid sequences can be produced by denovo solid-phase DNA synthesis or by PCR mutagenesis of an earlierprepared variant of the desired polynucleotide. Oligonucleotide-mediatedmutagenesis is a preferred method for preparing substitution, deletionand insertion variants of target polypeptide DNA. See Adelman et al.,DNA 2: 183 (1983). Briefly, the target polypeptide DNA is altered byhybridizing an oligonucleotide encoding the desired mutation to asingle-stranded DNA template. After hybridization, a DNA polymerase isused to synthesize an entire second complementary strand of the templatethat incorporates the oligonucleotide primer, and encodes the selectedalteration in the target polypeptide DNA.

Selection of Constant Region.

The variable segments of humanized antibodies produced as describedsupra are typically linked to at least a portion of an immunoglobulinconstant region (Fc), typically that of a human immunoglobulin. Humanconstant region DNA sequences can be isolated in accordance withwell-known procedures from a variety of human cells, but preferablyimmortalized B-cells (see Kabat et al., supra, and WO 87/02671) (each ofwhich is incorporated by reference in its entirety). Ordinarily, theantibody will contain both light chain and heavy chain constant regions.The heavy chain constant region usually includes CH1, hinge, CH2, CH3,and CH4 regions.

The humanized antibodies include antibodies having all types of constantregions, including IgM, IgG, IgD, IgA and IgE, and any isotype,including IgG1, IgG2, IgG3 and IgG4. When it is desired that thehumanized antibody exhibit cytotoxic activity, the constant domain isusually a complement-fixing constant domain and the class is typicallyIgG₁. When such cytotoxic activity is not desirable, the constant domainmay be of the IgG₂ class. The humanized antibody may comprise sequencesfrom more than one class or isotype.

Other Anti-VLA-4 Antibodies

Other anti-VLA-4 antibodies include but are not limited to HP1/2,HP-2/1, HP2/4, L25, and P4C2. These antibodies may also be administeredin an effective amount to diagnose and/or treat imflammatory bowelconditions as one skilled in the art as discussed herein and asgenerally known in the art would readily appreciate.

Frequently, monoclonal antibodies created in mice are later humanized toavoid the human anti-mouse antibody (HAMA) immune response in a humansubject injected with a mouse antibody. This occurs by CDR grafting orreshaping. Thus, typically the antibodies are first mouse monoclonalantibodies that through CDR grafting or reshaping become humanized, asdiscussed above for the 21.6 antibody.

Specifically, the humanized antibodies have specificity for VLA-4 andhave the ability to diagnose and/or treat imflammatory bowel conditions.These antibodies are derived from sources (e.g., mouse typically) thatat least one or more of the complementarity determining regions (CDRs)of the variable domains are derived from a donor non-human anti-VLA-4antibody, and in which there may or may not have been minimal alterationof the acceptor antibody heavy and/or light variable framework region inorder to retain donor antibody binding specificity. Preferably, theantigen binding regions of the CDR-grafted heavy chain variable domaincomprise the CDRs corresponding to positions 31-35 (CDR1), 50-65 (CDR2)and 95-102 (CDR3). In a preferred embodiment, the heavy chain furtherincludes non-human residues at framework positions 27-30 (Kabatnumbering). The heavy chain can further include non-human residues atframework position 75 (Kabat numbering). The heavy chain can furtherinclude non-human residues at framework position(s) 77-79 or 66-67 and69-71 or 84-85 or 38 and 40 or 24 (Kabat numbering). Preferably, theantigen binding regions of the CDR-grafted light chain variable domaincomprise CDRs corresponding to positions 24-34 (CDR1), 50-56 (CDR2) and89-97 (CDR3). In a preferred embodiment, the light chain furtherincludes non-human residues at framework positions 60 and 67 (Kabatnumbering). These residue designations are numbered according to theKabat numbering (Kabat et al., 5^(th) ed. 4 vol. SEQUENCES OF PROTEINSOF IMMUNOLOGICAL INTEREST, U.S. Department of Health Human Services,NIH, USA (1991)).

Synthesis and Humanization of Mouse Antibody HP 1/2. HP 1/2 is anotherantibody that is directed against VLA-4. The method of preparing ahumanized version of this antibody for use in human subjects isdescribed herein and is further described in U.S. Pat. No. 6,602,503assigned to Biogen, Inc., and hereby incorporated by reference in itsentirety. The sequences of the humanized antibodies are provided asfollows. The HP 1/2 V_(H) DNA sequence and its translated amino acidsequence are: 5′-gtc aaa ctg cag cag tct ggg gca gag ctt gtg aag cca ggggcc tca 48  N-Val Lys Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro GlyAla Ser    1               5                   10                  15   gtc aag ttg ttc tgc aca gct tct ggc ttc aac att aaa gac acc tat  96   Val Lys Leu Phe Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Thr Tyr                20                  25                  30    atg cactgg gtg aag cag agg cct caa cag ggc ctg gag tgg att gga 144    Met HisTrp Val Lys Gln Arg Pro Gln Gln Gly Leu Glu Trp Ile Gly            35                  40                  45    agg att gatcct gcg agt ggc gat act aaa tat gac ccg aag ttc cag 192    Arg Ile AspPro Ala Ser Gly Asp Thr Lys Tyr Asp Pro Lys Phe Gln        50                  55                  60    gtc aag gcc actatt aca gcg gac acg tcc tcc aac aca gcc tgg ctg 240    Val Lys Ala ThrIle Thr Ala Asp Thr Ser Ser Asn Thr Ala Trp Leu    65                  70                  75                  80   cag ctc agc agc ctg aca tct gag gac act gcc gtc tac tac tgt gca 288   Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys Ala                    85                  90                  95    gacgga atg tgg gta tca acg gga tat gct ctg gac ttc tgg ggc caa 336    AspGly Met Trp Val Ser Thr Gly Tyr Ala Leu Asp Phe Trp Gly Gln               100                 105                 110    ggg accacg gtc acc gtc tcc tca-3′ 360    Gly Thr Thr Val Thr Val Ser Ser-C           115                 120

A comparison between HP½ V_(H) the two sequences and a consensussequence of family IIC revealed that the only unusual residues are atamino acid positions 80, 98 and 121 (i.e., 79, 94 and 121 in Kabatnumbering). Although Tyr-80 is invariant in subgroup IIC other sequencedmurine V_(H) regions have other aromatic amino acids at this position,although none have Trp. The majority of human and murine V_(H)s have anarginine residue at Kabat position 94. The presence of Asp-94 in HP½V_(H) is extremely rare; there is only one reported example of anegatively charged residue at this position. Proline at Kabat position113 is also unusual but is unlikely to be important in the conformationof the CDRs because of its distance from them. The amino acids making upCDR1 have been found in three other sequenced murine V_(H) regions.However, CDR2 and CDR3 are unique to HP 1/2 and are not found in anyother reported murine V_(H).

The HP½ V_(K) DNA sequence and its translated amino acid sequence are asfollows: 5′-agt att gtg atg acc cag act ccc aaa ttc ctg ctt gtt tca gcagga  48  N-Ser Ile Val Met Thr Gln Thr Pro Lys Phe Leu Leu Val Ser AlaGly     1               5                   10                  15   gac agg gtt acc ata acc tgc aag gcc agt cag agt gtg act aat gat  96   Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Ser Val Thr Asn Asp                20                  25                  30    gta gcttgg tac caa cag aag cca ggg cag tct cct aaa ctg ctg ata 144    Val AlaTrp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile            35                  40                  45    tat tat gcatcc aat cgc tac act gga gtc cct gat cgc ttc act ggc 192    Tyr Tyr AlaSer Asn Arg Tyr Thr Gly Val Pro Asp Arg Phe Thr Gly        50                  55                  60    agt gga tat gggacg gat ttc act ttc acc atc agc act gtg cag gct 240    Ser Gly Tyr GlyThr Asp Phe Thr Phe Thr Ile Ser Thr Val Gln Ala    65                  70                  75                  80   gaa gac ctg gca gtt tat ttc tgt cag cag gat tat agc tct ccg tac 288   Glu Asp Leu Ala Val Tyr Phe Cys Gln Gln Asp Tyr Ser Ser Pro Tyr                    85                  90                  95    acgttc gga ggg ggg acc aag ctg gag atc-3′ 318    Thr Phe Gly Gly Gly ThrLys Leu Glu Ile-C                100                 105

HP½ V_(K) is a member of Kabat family V (Kabat et al., 5^(th) ed., 4vol., SEQUENCES OF PROTEINS OF IMMUNOLOGICAL INTEREST, U.S. Departmentof Health Human Services (1991)) and has no unusual residues. The aminoacids of CDR1 and CDR3 are unique. The amino acids making up CDR2 havebeen reported in one other murine V_(K).

Design of a CDR-grafted Anti-VLA-4 Antibody. To design a CDR-graftedanti-VLA-4 antibody, it was necessary to determine which residues ofmurine HP½ comprise the CDRs of the light and heavy chains. Threeregions of hypervariability amid the less variable framework sequencesare found on both light and heavy chains (Wu and Kabat, J. Exp. Med.132: 211-250 (1970); Kabat et al., (1991)). Inmost cases thesehypervariable regions correspond to, but may extend beyond, the CDR.CDRs of murine HP½ were elucidated in accordance with Kabat et al.,(1991) by alignment with other V_(H) and V_(K) sequences. The CDRs ofmurine HP½ V_(H) were identified and correspond to the residuesidentified in the humanized V_(H) sequences as follows: CDR1 AA₃₁-AA₃₅CDR2 AA₅₀-AA₆₆ CDR3 AA₉₉-AA₁₁₀

These correspond to AA₃₁-AA₃₅, AA₅₀-AA₆₅, and AA₉₅-AA₁₀₂, respectively,in Kabat numbering. The CDRs of murine HP½ V_(K) were identified andcorrespond to the residues identified in the humanized V_(K) sequencesas follows: CDR1 AA₂₄-AA₃₄ CDR2 AA₅₀-AA₅₆ CDR3 AA₈₉-AA₉₇These correspond to the same numbered amino acids in Kabat numbering.Thus, only the boundaries of the V_(K), but not V_(H), CDRs correspondedto the Kabat CDR residues. The human frameworks chosen to accept the HP½(donor) CDRs were NEWM and RE1 for the heavy and light chains,respectively. The NEWM and the RE1 sequences have been published inKabat et al., (1991).

The DNA and corresponding amino acid sequence of the humanized heavychain variable region of the humanized HP½ antibody is: 5′-atg gac tggacc tgg agg gtc ttc tgc ttg ctg gct gta gca cca ggt 48  N-Met Asp TrpThr Trp Arg Val Phe Cys Leu Leu Ala Val Ala Pro Gly    1               5                   10                  15    gcccac tcc cag gtc caa ctg cag gag tcc ggt gct gaa gtt gtt aaa 96    AlaHis Ser Gln Val Gln Leu Gln Glu Ser Gly Ala Glu Val Val Lys                20                  25                  30    ccg ggttcc tcc gtt aaa ctg tcc tgc aaa gct tcc ggt ttc aac atc 144    Pro GlySer Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Phe Asn Ile            35                  40                  45    aaa gac acctac atg cac tgg gtt aaa cag cgt ccg ggt cag ggt ctg 192    Lys Asp ThrTyr Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu        50                  55                  60    gaa tgg atc ggtcgt atc gac ccg gct tcc ggt gac acc aaa tac gac 240    Glu Trp Ile GlyArg Ile Asp Pro Ala Ser Gly Asp Thr Lys Tyr Asp    65                  70                  75                  80   ccg aaa ttc cag gtt aaa gct acc atc acc gct gac gaa tcc acc tcc 288   Pro Lys Phe Gln Val Lys Ala Thr Ile Thr Ala Asp Glu Ser Thr Ser                    85                  90                  95    accgct tac ctg gaa ctg tcc tcc ctg cgt tcc gaa gac acc gct gtt 336    ThrAla Tyr Leu Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val               100                 105                 110    tac tactgc gct gac ggt atg tgg gtt tcc acc ggt tac gct ctg gac 384    Tyr TyrCys Ala Asp Gly Met Trp Val Ser Thr Gly Tyr Ala Leu Asp           115                 120                 125    ttc tgg ggtcag ggt acc acg gtc acc gtc tcc tca ggt gag tcc-3′ 429    Phe Trp GlyGln Gly Thr Thr Val Thr Val Ser Ser Gly Glu Ser-C       130                 135                 140

The DNA and corresponding amino acid sequence of the humanized lightchain variable region of the humanized HP½ antibody: 5′-atg ggt tgg tcctgc atc atc ctg ttc ctg gtt gct acc gct acc ggt  48  N-Met Gly Trp SerCys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly    1               5                   10                  15    gttcac tcc atc gtt atg acc cag tcc ccg gac tcc ctg gct gtt tcc  96    ValHis Ser Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser                20                  25                  30    ctg ggtgaa cgt gtt acc atc aac tgc aaa gct tcc cag tcc gtt acc 144    Leu GlyGlu Arg Val Thr Ile Asn Cys Lys Ala Ser Gln Ser Val Thr            35                  40                  45    aac gac gttgct tgg tac cag cag aaa ccg ggt cag tcc ccg aaa ctg 192    Asn Asp ValAla Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu        50                  55                  60    ctg atc tac tacgct tcc aac cgt tac acc ggt gtt ccg gac cgt ttc 240    Leu Ile Tyr TyrAla Ser Asn Arg Tyr Thr Gly Val Pro Asp Arg Phe    65                  70                  75                  80   tcc ggt tcc ggt tac ggt acc gac ttc acc ttc acc atc tcc tcc gtt 288   Ser Gly Ser Gly Tyr Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Val                    85                  90                  95    caggct gaa gac gtt gct gtt tac tac tgc cag cag gac tac tcc tcc 336    GlnAla Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Asp Tyr Ser Ser               100                 105                 110    ccg tacacc ttc ggt ggt ggt acc aaa ctg gag atc taa ggatcctc-3′ 383    Pro TyrThr Phe Gly Gly Gly Thr Lys Leu Glu Ile-C           115                 120

In addition to the above humanized HP½ antibody light and heavy chains,other acceptor heavy and light chains regions can also be utilized forinsertion of the donor HP½ regions. All the following constructs containSer-75 (Kabat numbering). The STAW construct further contains Gln to Thrat position 77, Phe to Ala at position 78, and Ser to Trp at position 79(Kabat numbering). The V_(H) DNA sequence and its translated amino acidsequence are set forth below: 5′-atg gac tgg acc tgg agg gtc ttc tgc ttgctg gct gta gca cca ggt  48  N-Met Asp Trp Thr Trp Arg Val Phe Cys LeuLeu Ala Val Ala Pro Gly    1               5                   10                  15    gcccac tcc cag gtc caa ctg cag gag agc ggt cca ggt ctt gtg aga  96    AlaHis Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg                20                  25                  30    cct agccag acc ctg agc ctg acc tgc acc gtg tct ggc ttc aac att 144    Pro SerGln Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Phe Asn Ile            35                  40                  45    aaa gac acctat atg cac tgg gtg aga cag cca cct gga cga ggt ctt 192    Lys Asp ThrTyr Met His Trp Val Arg Gln Pro Pro Gly Arg Gly Leu        50                  55                  60    gag tgg att ggaagg att gat cct gcg agt ggc gat act aaa tat gac 240    Glu Trp Ile GlyArg Ile Asp Pro Ala Ser Gly Asp Thr Lys Tyr Asp    65                  70                  75                  80   ccg aag ttc cag gtc aga gtg aca atg ctg gta gac acc agc agc aac 288   Pro Lys Phe Gln Val Arg Val Thr Met Leu Val Asp Thr Ser Ser Asn                    85                  90                  95    acagcc tgg ctg aga ctc agc agc gtg aca gcc gcc gac acc gcg gtc 336    ThrAla Trp Leu Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val               100                 105                 110    tat tattgt gca gac gga atg tgg gta tca acg gga tat gct ctg gac 384    Tyr TyrCys Ala Asp Gly Met Trp Val Ser Thr Gly Tyr Ala Leu Asp           115                 120                 125    ttc tgg ggccaa ggg acc acg gtc acc gtc tcc tca ggt gag tcc-3′ 429    Phe Trp GlyGln Gly Thr Thr Val Thr Val Ser Ser Gly Glu Ser-C       130                 135                 140

The KAITAS construct contains the additional changes of Arg to Lys(position 66), Val to Ala (position 67), Met to Ile (position 69), Leuto Thr (position 70) and Val to Ala (position 71) (Kabat numbering. TheKAITAS V_(H) DNA sequence and its translated amino acid sequence are setforth below: 5′-atg gac tgg acc tgg agg gtc ttc tgc ttg ctg gct gta gcacca ggt  48  N-Met Asp Trp Thr Trp Arg Val Phe Cys Leu Leu Ala Val AlaPro Gly     1               5                   10                  15   gcc cac tcc cag gtc caa ctg cag gag agc ggt cca ggt ctt gtg aga  96   Ala His Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg                20                  25                  30    cct agccag acc ctg agc ctg acc tgc acc gtg tct ggc ttc aac att 144    Pro SerGln Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Phe Asn Ile            35                  40                  45    aaa gac acctat atg cac tgg gtg aga cag cca cct gga cga ggt ctt 192    Lys Asp ThrTyr Met His Trp Val Arg Gln Pro Pro Gly Arg Gly Leu        50                  55                  60    gag tgg att ggaagg att gat cct gcg agt ggc gat act aaa tat gac 240    Glu Trp Ile GlyArg Ile Asp Pro Ala Ser Gly Asp Thr Lys Tyr Asp    65                  70                  75                  80   ccg aag ttc cag gtc aaa gcg aca att acg gca gac acc agc agc aac 288   Pro Lys Phe Gln Val Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn                    85                  90                  95    cagttc agc ctg aga ctc agc agc gtg aca gcc gcc gac acc gcg gtc 336    GlnPhe Ser Leu Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val               100                 105                 110    tat tattgt gca gac gga atg tgg gta tca acg gga tat gct ctg gac 384    Tyr TyrCys Ala Asp Gly Met Trp Val Ser Thr Gly Tyr Ala Leu Asp           115                 120                 125    ttc tgg ggccaa ggg acc acg gtc acc gtc tcc tca ggt gag tcc-3′ 429    Phe Trp GlyGln Gly Thr Thr Val Thr Val Ser Ser Gly Glu Ser-C       130                 135                 140

The SSE construct comprises the additional changes of Ala to Ser(position 84) and Ala to Glu (position 85) (Kabat numbering). The SSEV_(H) DNA sequence and its translated amino acid sequence are set forthbelow: 5′-cag gtc caa ctg cag gag agc ggt cca ggt ctt gtg aga cct agccag  48  N-Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro SerGln     1               5                   10                  15   acc ctg agc ctg acc tgc acc gtg tct ggc ttc aac att aaa gac acc 96   Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Phe Asn Ile Lys Asp Thr                20                  25                  30    tat atgcac tgg gtg aga cag cca cct gga cga ggt ctt gag tgg att 144    Tyr MetHis Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile            35                  40                  45    gga agg attgat cct gcg agt ggc gat act aaa tat gac ccg aag ttc 192    Gly Arg IleAsp Pro Ala Ser Gly Asp Thr Lys Tyr Asp Pro Lys Phe        50                  55                  60    cag gtc aga gtgaca atg ctg gta gac acc agc agc aac cag ttc agc 240    Gln Val Arg ValThr Met Leu Val Asp Thr Ser Ser Asn Gln Phe Ser    65                  70                  75                  80   ctg aga ctc agc agc gtg aca tct gag gac acc gcg gtc tat tat tgt 288   Leu Arg Leu Ser Ser Val Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys                    85                  90                  95    gcagac gga atg tgg gta tca acg gga tat gct ctg gac ttc tgg ggc 336    AlaAsp Gly Met Trp Val Ser Thr Gly Tyr Ala Leu Asp Phe Trp Gly               100                 105                 110    caa gggacc acg gtc acc gtc tcc tca ggt gag tcc-3′ 372    Gln Gly Thr Thr ValThr Val Ser Ser Gly Glu Ser-C            115                 120

The KRS construct comprises the additional changes of Arg to Lys(position 38) and Pro to Arg (position 40) (Kabat numbering). The KRSV_(H) DNA sequence and its translated amino acid sequence are set forthbelow: 5′-atg gac tgg acc tgg agg gtc ttc tgc ttg ctg gct gta gca ccaggt  48  N-Met Asp Trp Thr Trp Arg Val Phe Cys Leu Leu Ala Val Ala ProGly     1               5                   10                  15   gcc cac tcc cag gtc caa ctg cag gag agc ggt cca ggt ctt gtg aga  96   Ala His Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg                20                  25                  30    cct agccag acc ctg agc ctg acc tgc acc gtg tct ggc ttc aac att 144    Pro SerGln Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Phe Asn Ile            35                  40                  45    aaa gac acctat atg cac tgg gtg aaa cag cga cct gga cga ggt ctt 192    Lys Asp ThrTyr Met His Trp Val Lys Gln Arg Pro Gly Arg Gly Leu        50                  55                  60    gag tgg att ggaagg att gat cct gcg agt ggc gat act aaa tat gac 240    Glu Trp Ile GlyArg Ile Asp Pro Ala Ser Gly Asp Thr Lys Tyr Asp    65                  70                  75                  80   ccg aag ttc cag gtc aga gtg aca atg ctg gta gac acc agc agc aac 288   Pro Lys Phe Gln Val Arg Val Thr Met Leu Val Asp Thr Ser Ser Asn                    85                  90                  95    cagttc agc ctg aga ctc agc agc gtg aca gcc gcc gac acc gcg gtc 336    GlnPhe Ser Leu Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val               100                 105                 110    tat tattgt gca gac gga atg tgg gta tca acg gga tat gct ctg gac 384    Tyr TyrCys Ala Asp Gly Met Trp Val Ser Thr Gly Tyr Ala Leu Asp           115                 120                 125    ttc tgg ggccaa ggg acc acg gtc acc gtc tcc tca ggt gag tcc-3′ 429    Phe Trp GlyGln Gly Thr Thr Val Thr Val Ser Ser Gly Glu Ser-C       130                 135                 140

The AS construct comprises the change Val to Ala at position 24 (Kabatnumbering). The AS V_(H) DNA sequence and its translated amino acidsequence are: 5′-atg gac tgg acc tgg agg gtc ttc tgc ttg ctg gct gta gcacca ggt  48  N-Met Asp Trp Thr Trp Arg Val Phe Cys Leu Leu Ala Val AlaPro Gly     1               5                   10                  15   gcc cac tcc cag gtc caa ctg cag gag agc ggt cca ggt ctt gtg aga 96   Ala His Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg                20                  25                  30    cct agccag acc ctg agc ctg acc tgc acc gcg tct ggc ttc aac att 144    Pro SerGln Thr Leu Ser Leu Thr Cys Thr Ala Ser Gly Phe Asn Ile            35                  40                  45    aaa gac acctat atg cac tgg gtg aga cag cca cct gga cga ggt ctt 192    Lys Asp ThrTyr Met His Trp Val Arg Gln Pro Pro Gly Arg Gly Leu        50                  55                  60    gag tgg att ggaagg att gat cct gcg agt ggc gat act aaa tat gac 240    Glu Trp Ile GlyArg Ile Asp Pro Ala Ser Gly Asp Thr Lys Tyr Asp    65                  70                  75                  80   ccg aag ttc cag gtc aga gtg aca atg ctg gta gac acc agc agc aac 288   Pro Lys Phe Gln Val Arg Val Thr Met Leu Val Asp Thr Ser Ser Asn                    85                  90                  95    cagttc agc ctg aga ctc agc agc gtg aca gcc gcc gac acc gcg gtc 336    GlnPhe Ser Leu Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val               100                 105                 110    tat tattgt gca gac gga atg tgg gta tca acg gga tat gct ctg gac 384    Tyr TyrCys Ala Asp Gly Met Trp Val Ser Thr Gly Tyr Ala Leu Asp           115                 120                 125    ttc tgg ggccaa ggg acc acg gtc acc gtc tcc tca ggt gag tcc-3′ 429    Phe Trp GlyGln Gly Thr Thr Val Thr Val Ser Ser Gly Glu Ser-C       130                 135                 140

The humanized light chain generally requires few, if any, modifications.However, in the preparation of humanized anti-VLA-4 antibodies, severalempirical changes did improve the immunological activity of the antibodytowards its ligand. For example, the humanized heavy chain with the Sermutation with the murine light chain was about 2:5 fold lower potencythan murine HP½. The same humanized heavy chain with a humanized lightchain was about 4-fold lower potency.

A humanized V_(K) construct (VK1) comprises a Ser to Asp substitution atposition 60, and a Ser for a Tyr at position 67. The DNA sequence andits translated amino acid sequence are set forth below: 5′-atg ggt tggtcc tgc atc atc ctg ttc ctg gtt gct acc gct acc ggt  48  N-Met Gly TrpSer Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly    1               5                   10                  15    gttcac tcc gac atc cag ctg acc cag agc cca agc agc ctg agc gcc  96    ValHis Ser Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala                20                  25                  30    agc gtgggt gac aga gtg acc atc acc tgt aag gcc agt cag agt gtg 144    Ser ValGly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Ser Val            35                  40                  45    act aat gatgta gct tgg tac cag cag aag cca ggt aag gct cca aag 192    Thr Asn AspVal Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys        50                  55                  60    ctg ctg atc tactat gca tcc aat cgc tac act ggt gtg cca agc aga 240    Leu Leu Ile TyrTyr Ala Ser Asn Arg Tyr Thr Gly Val Pro Ser Arg    65                  70                  75                  80   ttc agc ggt agc ggt agc ggt acc gac ttc acc ttc acc atc agc agc 288   Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser                    85                  90                  95    ctccag cca gag gac atc gcc acc tac tac tgc cag cag gat tat agc 336    LeuGln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Asp Tyr Ser               100                 105                 110    tct ccgtac acg ttc ggc caa ggg acc aag gtg gaa atc aaa cgt aag tg-3′ 386    SerPro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Lys-C           115                 120                 125

Another V_(K) construct (i.e., VK2) has the DQMDY sequences of theoriginal RE1 framework restored. The DNA and corresponding amino acidsequence are provided below: 5′-atg ggt tgg tcc tgc atc atc ctg ttc ctggtt gct acc gct acc ggt  48  N-Met Gly Trp Ser Cys Ile Ile Leu Phe LeuVal Ala Thr Ala Thr Gly    1               5                   10                  15    gtccac tcc agc atc gtg atg acc cag agc cca agc agc ctg agc gcc  96    ValHis Ser Ser Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ser Ala                20                  25                  30    agc gtgggt gac aga gtg acc atc acc tgt aag gcc agt cag agt gtg 144    Ser ValGly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Ser Val            35                  40                  45    act aat gatgta gct tgg tac cag cag aag cca ggt aag gct cca aag 192    Thr Asn AspVal Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys        50                  55                  60    ctg ctg atc tactat gca tcc aat cgc tac act ggt gtg cca gat aga 240    Leu Leu Ile TyrTyr Ala Ser Asn Arg Tyr Thr Gly Val Pro Asp Arg    65                  70                  75                  80   ttc agc ggt agc ggt tat ggt acc gac ttc acc ttc acc atc agc agc 288   Phe Ser Gly Ser Gly Tyr Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser                    85                  90                  95    ctccag cca gag gac atc gcc acc tac tac tgc cag cag gat tat agc 336    LeuGln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Asp Tyr Ser               100                 105                 110    tct ccgtac acg ttc ggc caa ggg acc aag gtg gaa atc aaa cgt aag tg-3′ 386 SerPro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Lys-C        115                 120                 125

A third V_(K) construct is VK3 has SVM versus DQM in the amino terminusand two other residue changes. The DNA and corresponding amino acidsequence are: 5′-atg ggt tgg tcc tgc atc atc ctg ttc ctg gtt gct acc gctacc ggt  48  N-Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr AlaThr Gly     1               5                   10                  15   gtc cac tcc gac atc cag atg acc cag agc cca agc agc ctg agc gcc  96   Val His Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala                20                  25                  30    agc gtgggt gac aga gtg acc atc acc tgt aag gcc agt cag agt gtg 144    Ser ValGly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Ser Val            35                  40                  45    act aat gatgta gct tgg tac cag cag aag cca ggt aag gct cca aag 192    Thr Asn AspVal Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys        50                  55                  60    ctg ctg atc tactat gca tcc aat cgc tac act ggt gtg cca gat aga 240    Leu Leu Ile TyrTyr Ala Ser Asn Arg Tyr Thr Gly Val Pro Asp Arg    65                  70                  75                  80   ttc agc ggt agc ggt tat ggt acc gac ttc acc ttc acc atc agc agc 288   Phe Ser Gly Ser Gly Tyr Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser                    85                  90                  95    ctccag cca gag gac atc gcc acc tac tac tgc cag cag gat tat agc 336    LeuGln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Asp Tyr Ser               100                 105                 110    tct ccgtac acg ttc ggc caa ggg acc aag gtg gaa atc aaa cgt aag tg-3′ 386    SerPro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Lys-C           115                 120                 125

Details regarding how each of these light and heavy chain sequences wereprepared are provided in U.S. Pat. No. 6,602,503, which is herebyincorporated by reference in its entirety for all puposes. Variouscombinations of the above light and heavy chains can be prepared basedon computer modeling as known in the art.

Additional antibodies that recognize and bind to α₄ integrin are knownin the art. These include but are not limited to GG5/3 (Keszthelyi etal., Neurology 47(4): 1053-1059 (1996)), FW3-218-1 (ATCC No.: HB-261; anIgG2b antibody against sheep α₄ integrin), and R1-2 (ATCC No.: HB-227;IgG2b antibody developed in Rattus norvegicus). Whether the antibodiesare developed in mouse or other animals, each of the sequences can begenetically engineered such that they are humanized based on what isknown in the art and with the aid of computer modeling. The anti-α₄integrin humanized antibodies can then be assessed for their ability todiagnose and/or treat imflammatory bowel conditions on the in vitro andin vivo assays disclosed herein.

Antibody Fragments. Also contemplated for use in treating rheumatoidarthritis are antibody fragments of antibodies that bind to anti-alpha4or VCAM-1 such that they inhibit VLA-4 and VCAM-1 interaction. Antibodyfragments include Fab, F(ab′)₂, scFv and Fv fragments which can be usedin the compositions disclosed herein.

The term “Fab fragment” as used herein refers to a partial antibodymolecule containing a single antigen-binding region, which consists of aportion of both the heavy and light chains of the molecule.

The term “F(ab′)₂ fragment” as used herein refers to a partial antibodymolecule containing both antigen binding regions, and which consists ofthe light chains and a portion of the heavy chains of the molecule.

The term “Fv fragment” as used herein refers to the portion of theantibody molecule involved in antigen recognition and binding.

The term “scFv” as used herein refers to single chain Fv (scFv)fragments. These scFv fragments are recombinant antibody derivativesthat consist only of the variable domains of antibody heavy and lightchains connected by a flexible linker. scFv antibody fragments comprisethe V_(H) and V_(L) domains of antibody, wherein these domains arepresent in a single polypeptide chain. Generally, the Fv polypeptidefurther comprises a polypeptide linker between the V_(H) and V_(L)domains which enables the scFv to form the desired structure for antigenbinding. For a review of scFv see Pluckthun in The Pharmacology ofMonoclonal Antibodies, vol. 113, 269-315 (Rosenburg and Moore eds.,Springer-Verlag, New York 1994).

Also included in antibody fragments are diabodies. The term “diabodies”refers to small antibody fragments with two antigen-binding sites, whichfragments comprise a heavy chain variable domain (V_(H)) connected to alight chain variable domain (V_(L)) in the same polypeptide chain(V_(H)−V_(L)). By using a linker that is too short to allow pairingbetween the two domains on the same chain, the domains are forced topair with the complementary domains of another chain and create twoantigen-binding sites. Diabodies are described more fully in, forexample, EP 404,097; WO 93/11161; and Hollinger et al., 1993 Proc. Natl.Acad. Sci. USA 90: 6444-8.

Antibody fragments also include linear antibodies. The expression“linear antibodies” when used throughout this application refers to theantibodies described in, e.g., Zapata et al., 1995 Protein Eng. 8(10):1057-62. Briefly, these antibodies comprise a pair of tandem Fd segments(V_(H)-C_(H)1-V_(H)-C_(H)1), which form a pair of antigen bindingregions. Linear antibodies can be bispecific or monospecific.

Papain digestion of antibodies produces two identical antigen bindingfragments, called “Fab” fragments, each with a single antigen bindingsite, and a residual “Fc” fragment, whose name reflects its ability tocrystallize readily. Pepsin treatment yields an F(ab′)₂ fragment thathas two antigen combining sites and is still capable of cross-linkingantigen.

Antibody Purification. When using recombinant techniques, the antibodycan be produced intracellularly, in the periplasmic space, or directlysecreted into the medium. If the antibody is produced intracellularly,as a first step, the particulate debris, either host cells or lysedfragments, is removed, for example, by centrifugation orultrafiltration. Carter et al., Bio/Technology 10: 163-7 (1992) describea procedure for isolating antibodies, which are secreted to theperiplasmic space of E. coli. Briefly, cell paste is thawed in thepresence of sodium acetate (pH 3.5), EDTA, andphenylmethylsulfonylfluoride (PMSF) over.about 30 min. Cell debris canbe removed by centrifugation. In instances when the antibody is secretedinto the medium, supernatants from such expression systems are generallyfirst concentrated using a commercially available protein concentrationfilter, for example, an Amicon or Millipore Pellicon ultrafiltrationunit. A protease inhibitor such as PMSF may be included in any of theforegoing steps to inhibit proteolysis and antibiotics may be includedto prevent the growth of adventitious contaminants.

The antibody composition prepared from the cells is preferably subjectedto at least one purification step prior to LPHIC. Examples of suitablepurification steps include hydroxylapatite chromatography, gelelectrophoresis, dialysis, and affinity chromatography, with affinitychromatography being the preferred purification technique. Thesuitability of protein A as an affinity ligand depends on the speciesand isotype of any immunoglobulin Fc domain that is present in theantibody. Protein A can be used to purify antibodies that are based onhuman γ1, γ2, or γ4 heavy chains (Lindmark et al., 1983 J. Immunol.Meth. 62: 1-13). Protein G is recommended for all mouse isotypes and forhuman 73 (Guss et al., 1986 EMBO J. 5: 1567-75). The matrix to which theaffinity ligand is attached is most often agarose, but other matricesare available. Mechanically stable matrices such as controlled poreglass or poly(styrenedivinyl)benzene allow for faster flow rates andshorter processing times than can be achieved with agarose. Where theantibody comprises a C_(H)3 domain, the Bakerbond ABX™ resin (J. T.Baker, Phillipsburg, N.J.) is useful for purification. Other techniquesfor protein purification such as fractionation on an ion-exchangecolumn, ethanol precipitation, Reverse Phase HPLC, chromatography onsilica, chromatography on heparin SEPHAROSE™, chromatography on an anionor cation exchange resin (such as a polyaspartic acid column),chromatofocusing, SDS-PAGE, and ammonium sulfate precipitation are alsoavailable depending on the antibody to be recovered.

Following any preliminary purification step(s), the mixture comprisingthe antibody of interest and contaminant(s) is subjected to LPHIC.Often, the antibody composition to be purified will be present in abuffer from the previous purification step. However, it may be necessaryto add a buffer to the antibody composition prior to the LPHIC step.Many buffers are available and can be selected by routineexperimentation. The pH of the mixture comprising the antibody to bepurified and at least one contaminant in a loading buffer is adjusted toa pH of about 2.5-4.5 using either an acid or base, depending on thestarting pH. Preferably, the loading buffer has a low salt concentration(i.e., less than about 0.25 M salt).

The mixture is loaded on the HIC column. HIC columns normally comprise abase matrix (e.g., cross-linked agarose or synthetic copolymer material)to which hydrophobic ligands (e.g., alkyl or aryl groups) are coupled. Apreferred HIC column comprises an agarose resin substituted with phenylgroups (e.g., a Phenyl SEPHAROSE™ column). Many HIC coluimns areavailable commercially. Examples include, but are not limited to, PhenylSEPHAROSE 6 FAST FLOW™ column with low or high substitution (PharmaciaLKB Biotechnology, AB, Sweden); Phenyl SEPHAROSE™ High Performancecolumn (Pharmacia LKB Biotechnology, AB, Sweden); Octyl SEPHAROSE™ HighPerfonnance column (Pharmacia LKB Biotechnology, AB, Sweden); FRACTOGEL™EMD Propyl or FRACTOGEL™ EMD Phenyl columns (E. Merck, Germany);MACRO-PREP™ Methyl or MACRO-PREP™ t-Butyl Supports (Bio-Rad,California); WP HI-Propyl (C₃)™ column (J. T. Baker, New Jersey); andTOYOPEARL™ ether, phenyl or butyl columns (TosoHaas, Pa.).

The antibody is eluted from the column using an elution buffer, which isnormally the same as the loading buffer. The elution buffer can beselected using routine experimentation. The pH of the elution buffer isbetween about 2.5-4.5 and has a low salt concentration (i.e., less thanabout 0.25 M salt). It has been discovered that it is not necessary touse a salt gradient to elute the antibody of interest; the desiredproduct is recovered in the flow through fraction, which does not bindsignificantly to the column.

The LPHIC step provides a way to remove a correctly folded and disulfidebonded antibody from unwanted contaminants (e.g., incorrectly associatedlight and heavy fragments). In particular, the method provides a meansto substantially remove an impurity characterized-herein as a correctlyfolded antibody fragment whose light and heavy chains fail to associatethrough disulfide bonding.

Diagnostic or therapeutic formulations of the purified protein can bemade by providing the antibody composition in the form of aphysiologically acceptable carrier, examples of which are providedbelow.

To remove contaminants (e.g., unfolded antibody and incorrectlyassociated light and heavy fragments) from the HIC column so that it canbe re-used, a composition including urea (e.g., 6.0 M urea, 1% MESbuffer pH 6.0, 4 mM ammonium sulfate) can be flowed through the column.Other methods are known in the art.

Immunoglobulin Formulations. Antibodies and immunoglobulins having thedesired therapeutic effect may be administered in a physiologicallyacceptable carrier to a subject. The antibodies may be administered in avariety of ways including but not limited to parenteral administration,including subcutaneous, subdural, intravenous, intramuscular,intrathecal, intraperitoneal, intracerebral, intraarterial, orintralesional routes of administration, localized (e.g., surgicalapplication or surgical suppository), and pulmonary (e.g., aerosols,inhalation, or powder).

Depending upon the manner of introduction, the immunglobulins may beformulated in a variety of ways. The concentration of therapeuticallyactive immunoglobulin in the formulation (i.e., a formulation sufficientto inhibit rheumatoid arthritis) may vary from about 1 mg/ml to 1 g/ml.Preferably, the immunoglobulin composition, when administered to asubject in need thereof, reaches a blood level of immunoglobulin in thesubject of about 10 ng/ml or more.

Preferably, the immunoglobulin is formulated for parenteraladministration in a suitable inert carrier, such as a sterilephysiological saline solution. For example, the concentration ofimmunoglobulin in the carrier solution is typically between about 1-100mg/ml. The dose administered will be determined by route ofadministration. Preferred routes of administration include parenteral orintravenous administration.

According to one aspect of the invention, the immunoglobulins areadministered in combination with methotrexate, to treat, ameliorate, orpalliate the symptoms of rheumatoid arthritis. When administered incombination, the immunoglobulins may be administered in the sameformulation as the methotrexate, or in a separate formulation. Thecompounds may be administered prior to, following, or concurrently withthe methotrexate such that the benefitsd of the combination therapy areachieved. The calculation of appropriate dosages will be well within thepurvue of the skilled artisan. Standard doses of methotrexate for thetreatment of rheumatoid arthritis range from 2 mg to 20 mg per dose perweek. Dosages of the compounds are as set forth above. The methotrexatedosage may be administered as a single dose or as a divided dose. Once aresponse has been achieved, the dosage may be reduced if possible to thelowest effective dose. The maximum recommended dose is 20 mg/week.Preferably, methotrexate is administered orally or via injection.

According to an important feature of the invention, an immunoglobulinthat recognizes and binds to VLA-4 may be administered alone, or incombination with an anti-inflammatory agent, which is typically used totreat rheumatoid arthritis. Administration of anti-inflammatory agentscan occur prior to, concurrent with or after administration with theimmunoglobulin.

A therapeutically effective amount of an anti-alpha-4 integrin antibodyor immunoglobulin, e.g., Antegren™ (also known as natalizumab), can beestimated by comparison with established effective doses for knownantibodies, taken together with data obtained for Antegren™ in both invivo and in vitro models. As is known in the art, adjustments in thedose may be necessary due to immunoglobulin degeneration or metabolism,systemic versus localized delivery, as well as the age, body weight,general health, sex, diet, time of administration, drug interactions andthe severity of the condition of the subject to whom the immunoglobulinis administered. Such adjustments may be made and appropriate dosesdetermined by one of skill in the art through routine experimentation.

Therapeutic formulations of the immunoglobulin are prepared for storageby mixing the immunoglobulin having the desired degree of purity withoptional physiologically acceptable carriers, excipients, or stabilizers(Remington's Pharmaceutical Sciences, 16^(th) ed., A. Osol, Ed., 1980and more recent editions), in the form of lyophilized cake or aqueoussolutions. Acceptable immunoglobulin carriers, excipients or stabilizersare nontoxic, nontherapeutic and/or nonimmunogenic to recipients at thedosages and concentrations employed, and include buffers such asphosphate, citrate, and other organic acids; antioxidants includingascorbic acid; low molecular weight (less than about 10 residues)polypeptides; proteins, such as serum albumin, gelatin, orimmunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone;amino acids such as glycine, glutamine, asparagine, arginine or lysine;monosaccharides, disaccharides, and other carbohydrates includingglucose, mannose, or dextrins; chelating agents such as EDTA; sugaralcohols such as mannitol or sorbitol; salt-forming counterions such assodium; and/or nonionic surfactants such as Tween, Pluronics orpolyethylene glycol (PEG). Specific examples of carrier moleculesinclude but are not limited to glycosaminoglycans (e.g., heparinsulfate), hyaluronic acid, keratan-sulfate, chondroitin 4-sulfate,chondroitin 6-sulfate, heparan sulfate and dermatin sulfate, perlecanand pentopolysulfate.

Pharmaceutical compositions comprising immunoglobulins can also includeif desired, pharmaceutically acceptable, non-toxic carriers or diluents,which are vehicles commonly used to formulated pharmaceuticalcompositions for animal or human administration. The diluent is selectedso as not to affect the biological activity of the combination. Examplesinclude but are not limited to distilled water, physiologicalphosphate-buffered saline, Ringer's solutions, dextrose solution, andHank's solution.

The agents of the invention can be formulated into preparations forinjections by dissolving, suspending or emulsifying them in an aqueousor nonaqueous solvent, such as vegetable or other similar oils,synthetic aliphatic acid glycerides, esters of higher aliphatic acids orpropylene glycol. The formulations may also contain conventionaladditives, such as solubilizers, isotonic agents, suspending agents,emulsifying agents, stabilizers and preservatives.

The immunoglobulins may also be utilized in aerosol formulation to beadministered via inhalation or pulmonary delivery. The agents of thepresent invention can be formulated into pressurized acceptablepropellants such as dichlorodifluoromethane, propane, nitrogen and thelike.

The immunoglobulin also may be entrapped in microcapsules prepared, forexample, by coacervation techniques or by interfacial polymerization(e.g., hydroxymethylcellulose or gelatin-microcapsules andpoly-methylmethacylate microcapsules), in colloidal drug deliverysystems (e.g., liposomes, albumin microspheres, microemulsions,nano-particles and nanocapsules), or in macroemulsions. Such techniquesare disclosed in Remington's Pharmaceutical Sciences, supra.

The immunglobulin-to be used for in vivo administration must be sterile.This is readily accomplished by filtration through sterile filtrationmembranes, prior to or following lyophilization and reconstitution. Theimmunglobulin ordinarily will be stored in lyophilized form or insolution.

Therapeutic immunglobulin compositions generally are placed into acontainer having a sterile access port, for example, an intravenoussolution bag or vial having a stopper pierceable by a hypodermicinjection needle or similar sharp instrument.

Suitable examples of sustained-release preparations includesemipermeable matrices of solid hydrophobic polymers containing theprotein, which matrices are in the form of shaped articles, e.g., films,or microcapsules. Examples of sustained-release matrices includepolyesters, hydrogels (e.g., poly(2-hydroxyethyl-methacrylate) asdescribed by Langer et al., J. Biomed. Mater. Res. 15: 167-277 (1981)and Langer, Chem. Tech. 12: 98-105 (1982) or poly(vinylalcohol)),polylactides (U.S. Pat. No. 3,773,919), copolymers of L-glutamic acidand gamma ethyl-L-glutamate (Sidman et al., Biopolymers 22: 547-556,1983), non-degradable ethylene-vinyl acetate (Langer et al., supra),degradable lactic acid-glycolic acid copolymers such as the LUPRONDEPOT™ (i.e., injectable microspheres composed of lactic acid-glycolicacid copolymer and leuprolide acetate), and poly-D-(−)-3-hydroxybutyricacid (EP 133,988).

While polymers such as ethylene-vinyl acetate and lactic acid-glycolicacid enable release of molecules for over 100 days, certain hydrogelsrelease proteins for shorter time periods. When encapsulated antibodiesremain in the body for a long time, they may denature or aggregate as aresult of exposure to moisture at 37° C., resulting in a loss ofbiological activity and possible changes in immunogenicity. Rationalstrategies can be devised for immunglobulin stabilization depending onthe mechanism involved. For example, if the aggregation mechanism isdiscovered to be intermolecular S—S bond formation throughthio-disulfide interchange, stabilization may be achieved by modifyingsulflhydryl residues, lyophilizing from acidic solutions, controllingmoisture content, using appropriate additives, developing specificpolymer matrix compositions, and the like.

Sustained-release immunglobulin compositions also include liposomallyentrapped immunglobulin. Liposomes containing the immunglobulin areprepared by methods known per se. See, e.g., Epstein et al., Proc. Natl.Acad. Sci. USA 82: 3688-92 (1985); Hwang et al., Proc. Natl. Acad. Sci.USA 77: 4030-4 (1980); U.S. Pat. Nos. 4,485,045; 4,544,545; 6,139,869;and 6,027,726. Ordinarily, the liposomes are of the small (about 200 toabout 800 Angstroms), unilamellar type in which the lipid content isgreater than about 30 mole percent (mol. %) cholesterol; the selectedproportion being adjusted for the optimal immunglobulin therapy.

The immunoglobulins of this invention can be administered in a sustainedrelease form, for example a depot injection, implant preparation, orosmotic pump, which can be formulated in such a manner as to permit asustained release of the active ingredient. Implants for sustainedrelease formulations are well-known in the art. Implants are formulatedas microspheres, slabs, etc. with biodegradable or non-biodegradablepolymers. For example, polymers of lactic acid and/or glycolic acid forman erodible polymer that is well-tolerated by the host. The implant isplaced in proximity to the site of protein deposits (e.g., the site offormation of amyloid deposits associated with neurodegenerativedisorders), so that the local concentration of active agent is increasedat that site relative to the rest of the body.

In addition, immunoglobulins which prevent RA may be provided byadministering a polynucleotide encoding a whole or partial antibody(e.g., a single chain Fv) to a subject. The polynucleotide isadministered to a subject in an appropriate vehicle to allow theexpression of the immunoglobulin in the subject in a therapeuticallyeffective amount.

Certain agents of the invention, including antibodies and peptides, aresometimes administered in combination with an adjuvant. A variety ofadjuvants can be used in combination with an anti alpha-4 agent toelicit an immune response. Preferred adjuvants augment the intrinsicresponse to an agent without causing conformational changes in the agentthat affect the qualitative form of the response. Preferred adjuvantsinclude aluminum hydroxide and aluminum phosphate, 3 De-O-acylatedmonophosphoryl lipid A (MPL™) (see GB 2220211 (RIBI ImmunoChem ResearchInc., Hamilton, Mont., now part of Corixa). Stimulon™ QS-21 is atriterpene glycoside or saponin isolated from the bark of the QuillajaSaponaria Molina tree found in South America (see Kensil et al., inVaccine Design: The Subunit and Adjuvant Approach (eds. Powell & Newman,Plenum Press, New York, 1995); U.S. Pat. No. 5,057,540), (AquilaBioPharmaceuticals, Framingham, Mass.). Other adjuvants are oil in wateremulsions (such as squalene or peanut oil), optionally in combinationwith immune stimulants, such as monophosphoryl lipid A (see Stoute etal., 1997 N. Engl. J. Med. 336: 86-91). Another adjuvant is CpG (WO98/40100). Alternatively, an agent can be coupled to an adjuvant.However, such coupling should not substantially change the conformationof the desired alpha-4 epitope so as to affect the nature of the hostimmune response. Adjuvants can be administered as a component of atherapeutic composition with an active agent or can be administeredseparately, before, concurrently with, or after administration of thetherapeutic agent.

A preferred class of adjuvants for administration is aluminum salts(alum), such as aluminum hydroxide, aluminum phosphate, aluminumsulfate. Such adjuvants can be used with or without other specificimmunostimulating agents such as MPL or 3-DMP, QS-21, polymeric ormonomeric amino acids such as polyglutamic acid or polylysine. Anotherclass of adjuvants is oil-in-water emulsion formulations. Such adjuvantscan be used with or without other specific immunostimulating agents suchas muramyl peptides (e.g., N-acetylmuramyl-L-threonyl-D-isoglutamine(thr-MDP), N-acetyl-normuramyl-L-alanyl-D-isoglutamine (nor-MDP),N-acetylmuramyl-L-alanyl-D-isoglutaminyl-L-alanine-2-(1′-2′dipalmitoyl-sn-glycero-3-hydroxyphosphoryloxy)-ethylamine(MTP-PE),N-acetylglucsaminyl-N-acetylmuramyl-L-Al-D-isoglu-L-Ala-dipalmitoxypropylamide (DTP-DPP) theramide™, or other bacterial cell wallcomponents. Oil-in-water emulsions include (a) MF59 (WO 90/14837),containing 5% Squalene, 0.5% Tween 80, and 0.5% Span 85 (optionallycontaining various amounts of MTP-PE) formulated into submicronparticles using a microfluidizer such as Model 110Y microfluidizer(Microfluidics, Newton Mass.), (b) SAF, containing 10% Squalene, 0.4%Tween 80, 5% pluronic-blocked polymer L1 21, and thr-MDP, eithermicrofluidized into a submicron emulsion or vortexed to generate alarger particle size emulsion, and (c) Ribi™ adjuvant system (RAS),(Ribi Immunochem, Hamilton, Mont.) containing 2% squalene, 0.2% Tween80, and one or more bacterial cell wall components from the groupconsisting of monophosphoryl lipid A, trehalose dimycolate (TDM), andcell wall skeleton (CWS), preferably MPL+CWS (Detox™). Another class ofpreferred adjuvants is saponin adjuvants, such as Stimulon™ (QS-2 1;Aquila, Framingham, Mass.) or particles generated therefrom such asISCOMs (immunostimulating complexes) and ISCOMATRIX. Other adjuvantsinclude Incomplete Freund′s Adjuvant (IFA), cytokines, such asinterleukins (e.g., IL-1, IL-2, and IL-12), macrophage colonystimulating factor (M-CSF), and tumor necrosis factor (TNF). Suchadjuvants are generally available from commercial sources.

An adjuvant can be administered with an agent as a single composition,or can be administered before, concurrent with or after administrationof the agent. The agent and an adjuvant can be packaged and supplied inthe same vial or can be packaged in separate vials and mixed before use.The agent and adjuvant are typically packaged with a label indicatingthe intended therapeutic application. If the agent and adjuvant arepackaged separately, the packaging typically includes instructions formixing before use. The choice of an adjuvant and/or carrier depends onsuch factors as the stability of the formulation containing theadjuvant, the route of administration, the dosing schedule, and theefficacy of the adjuvant for the species being vaccinated. In humans, apreferred pharmaceutically acceptable adjuvant is one that has beenapproved for human administration by pertinent regulatory bodies.Examples of such preferred adjuvants for humans include alum, MPL andQS-21. Optionally, two or more different adjuvants can be usedsimultaneously. Preferred combinations include alum with MPL, alum withQS-21, MPL with QS-21, and alum, QS-21 and MPL together. Also,Incomplete Freund′s adjuvant can be used (Chang et al., Advanced DrugDelivery Reviews 32, 173-186 (1998)), optionally in combination with anyof alum, QS-21, and MPL and all combinations thereof.

Antegren™. Antegren™ is a humanized monoclonal antibody that hastherapeutic potential in that it prevents migration of inflammatorycells from blood vessels to sites of inflammation. AntegrenTm binds tocell surface receptors known as alpha-4-beta-1 (VLA-4) and alpha-4-beta7integrins. These receptors help white blood cells, particularly Tlymphocytes and eosinophils, move from the inside of blood vessels outinto surrounding tissues of the body at sites of inflammation, wherethese cells then participate in the inflammatory process. Antegren™blocks these receptors, thus preventing the cells from contributing tothe inflammatory response.

The typical method of administering Antegren™ is intravenous.Intravenous administration requires the final formulation to beisotonic. For example, a formulation of Antegren™, 5 mg/mL in 50 mML-histidine, 150 mM NaCl, pH 6.0 was initially chosen. During a Phase IIstudy, protein precipitation of the antibody was observed during thedilution and introduction of Antegren™ into the clinical dosingapparatus. Polysorbate 80 was introduced into the formulation to resolvethe observed protein precipitation.

The two factors that have been shown to accelerate the precipitation ofthe Antegren™ antibody are the presence of trace levels of silicone oiland denaturation at the air-liquid interface. The silicone oil wasintroduced into the product upon use of standard lubricatedpolypropylene syringes equipped with siliconized rubber stoppers. Theintroduction of the silicone oil is sufficient to cause discernibleantibody precipitation upon gentle agitation and room temperaturestorage. The aggregation and subsequent precipitation caused bydenaturation at the air-liquid interface has become more discernableproblematic with the drug being shipped to more clinical sites. Bothcauses of protein precipitation have been resolved by the addition ofpolysorbate 80 at a concentration of 0.02% (w/v).

The addition of polysorbate 80 to the formulation also overcomes theproblem of precipitating or aggregating antibody when preparingformulations with higher protein content. Initial work focused onagitation-induced aggregation at high protein concentrations, including50 mg/mL. By subjecting the material to agitation using a vortex-typemixer, aggregated species were detected by size exclusion-highperformance liquid chromatography (SEC-HPLC). This model identifiedpolysorbate 80 as an effective inhibitor of aggregation, while sucroseand other buffering components had little beneficial effect.

The effectiveness of the addition of 0.02% (w/v) polysorbate 80 inpreventing agitation-induced precipitation at a protein concentration of5 mg/mL was assessed following addition of 10 μL of a 10% polysorbate 80solution to vials of Antegren™ (Lot No. AN100226-0003). The vials wereshaken on their sides along with several vials of Antegren™ inFormulation #1 at 150 rotations per minute in a horizontal plane. Within3 hours of this treatment at room temperature, the vials of Formulation#1 were laden with particles and appeared turbid while the vials with0.02% (w/v) polysorbate 80 remained clear and free of particles.

The observed aggregation is presumed to be caused by the air-surfaceinterface, as vials completely filled with Antegren™ in the absence ofpolysorbate 80 were shaken for extended periods of time withoutadditional particle formation being induced.

An evaluation of the ability of 0.02% (w/v) polysorbate 80 to inhibitthe protein precipitation facilitated by trace levels of silicone wasconducted. A vial of Antegren™ (Lot No. AN100226-0003) was adjusted to0.02% (w/v) polysorbate 80 and drawn into a commercially available,lubricated 60 mL polypropylene syringe. The material was allowed tostand for several hours at room temperature. Visual inspection confirmedthat no precipitation was occurring. The material was then filteredthrough a 0.2 μm filter into a 5-mL vial and inspected and found to besubstantially free of particles after several days, while vials treatedin the same manner in the absence of polysorbate 80 (Formulation #1)were laden with particles.

Further descriptions of Antegren™ and procedures for preparing thishumanized monoclonal antibody are also described in U.S. Pat. No.5,840,299, which is herein incorporated by reference in its entirety.

Further descriptions of formulations for Antegren™ and procedures forpreparing such forumulations of this humanized monoclonal antibody arealso described in U.S. provisional Application Ser. No. 60/445,181,which is herein incorporated by reference in its entirety.

Agents and Small Molecule Alpha-4 Integrin Antogonist Compounds thatSelectively Bind to Alpha-4 Integrins

Various types of agents with the ability to bind to and inhibit alpha-4integrin can be used in the practice of the invention. Many such agentshave been identified and characterized, and specific agents aredescribed below. Given the teachings disclosed herein, it is well withinthe skill of one in the art to identify other agents that will be ableto inhibit the alpha-4-comprising integrin dimers in a manner thatbiologically mimics or is similar to the specifically described agents,and the present invention is intended to include the chronicadministration of such agents. As it is also contemplated to includecombinations of agents, discussion of agents other than small compoundsare also provided.

Drug Combinations. The anti-alpha-4 integrin agents (e.g., anti-alpha-4integrin antibodies and methotrexate and small compound alpha-4 integrinantagonists and methotrexate) can be combined with other compounds orcompositions used to treat, ameliorate or palliate symptoms associatedwith RA.

Dosage forms of the agents to be used in combination with the compoundsand compositions disclosed herein would vary depending on the subjectand drug combination being utilized.

The benefit of such combination therapies is that it may lessen theclass-specific and agent-specific side effects currently encounteredwith some of the drugs. Combinations of drugs that can lessen thequantity of a particular drug administered may reduce adverse sideeffects experienced by a patient.

When administered in combination, the small compound alpha-4-integrinantagonists may be administered in the same formulation as these othercompounds or compositions, or in a separate formulation. Whenadministered in combination, the anti-alpha-4-antibodies are generallyadministered in a separate formulation than the other compounds andcompositions. When administered in combinations, the anti-alpha-4 agentsmay be administered prior to, following, or concurrently with the othercompounds and compositions used to treat, ameliorate, or palliatesymptoms.

Chronic Administration Dosage Regimes

The chronic treatment regime of the present invention providesanti-alpha-4 agent at a level that will maintain sufficient receptorsaturation to treat rheumatoid arthritis in a patient in need of such.The methods of the invention entails administration once per every twoweeks or once a month to once every two months, with repeated dosingstaking place over a period of at least six months, and more preferablyfor a year or longer. The methods of the invention involve obtaining andmaintaining a receptor saturation level in a human patient of a dimercomprising alpha-4 integrin (e.g., VLA-4) in a range of from about 65%to 100%, more preferably between 75%, to 100%, and even more preferablybetween 80-100%. These receptor saturation levels are maintained atthese levels chronically (e.g., over a period of 6 months or so) toallow for continued suppression of pathological inflammation.

In a specific embodiment, the anti-alpha-4 agent is an antibody,preferably a humanized or human antibody, and the dosing is on a monthlybasis. In another specific embodiment, the anti-alpha-4 agent is acompound of Formula I-XXIX as provided above. Levels of receptorsaturation can be monitored to determine the efficacy of the dosingregime, and physiological markers measured to confirm the success of thedosage regime. As a confirmation, serum levels of the antibody can bemonitored to identify clearance of the antibody and to determine thepotential effect of half-life on the efficacy of the treatment.

The amount of agent administered in a dosage unit may depend on whetheradjuvant is also administered, with higher dosages generally beingrequired in the presence of adjuvant. For immunization with an agent ofthe invention, the dosage ranges from about 0.0001 to 100 mg/kg, andmore usually 0.01 to 5 mg/kg, of the host body weight. For exampledosages can be 1 mg/kg body weight or 10 mg/kg body weight. Dosage andfrequency vary depending on the half-life of the agent in the patient.The dosage and frequency of administration can vary depending on whetherthe treatment is prophylactic or therapeutic. For an antibodyadministration, each dosing injection is generally between 2.0 to 8.0mg/kg dosage. For a compound administration, each dosing injection isgenerally between 1.0 to 10.0 mg/kg dosage. In accordance with theteachings provided herein, effective dosages can be monitored byobtaining a fluid sample from a patient. For this, generally a bloodserum or cerebrospinal fluid sample it taken and integrin receptorsaturation is determined using methods well known in the art. Ideally, asample is taken prior to initial dosing; subsequent samples are takenand measured prior to and/or after each immunization. When adjuvant isbeing administered, the dosage level is increased in accordance with theparticular adjuvant and the level of immunogenicity of the anti-alpha-4agent. Doses for individual agents, selected in accordance with thepresent invention, are determined according to standard dosing methods,taken in conjunction with the teachings provided herein.

As an alternative to chronic administration comprised of repeatedindividual dosings, an anti-alpha-4 agent can be administered as asustained release formulation, provided the dosage is such that thelevels of receptor saturation remain sufficient to suppressinflammation. For example, controlled release systems can be used tochronically administer an anti-alpha-4 agent within the scope of thisinvention. Discussions of appropriate controlled release dosage formsmay be found in Lesczek Krowczynski, Extended-Release Dosage Forms, 1987(CRC Press, Inc.).

The various controlled release technologies cover a very broad spectrumof drug dosage forms. Controlled release technologies include, but arenot limited to physical systems and chemical systems. Physical systemsinclude, but not limited to, reservoir systems with rate-controllingmembranes, such as microencapsulation, macroencapsulation, and membranesystems; reservoir systems without rate-controlling membranes, such ashollow fibers, ultra microporous cellulose triacetate, and porouspolymeric substrates and foams; monolithic systems, including thosesystems physically dissolved in non-porous, polymeric, or elastomericmatrices (e.g., non-erodible, erodible, environmental agent ingression,and degradable), and materials physically dispersed in non-porous,polymeric, or elastomeric matrices (e.g., non-erodible, erodible,environmental agent ingression, and degradable); laminated structures,including reservoir layers chemically similar or dissimilar to outercontrol layers; and other physical methods, such as osmotic pumps, oradsorption onto ion-exchange resins.

Chemical systems include, but are not limited to, chemical erosion ofpolymer matrices (e.g., heterogeneous, or homogenous erosion), orbiological erosion of a polymer matrix (e.g., heterogeneous, orhomogeneous). Additional discussion of categories of systems forcontrolled release may be found in Agis F. Kydonieus, Controlled ReleaseTechnologies: Methods, Theory and Applications, 1980 (CRC Press, Inc.).

The methods of the invention can be used to treat a patient that isaffected with a disorder involving or arising from pathologicalinflammation, or to prophylactically treat a patient at risk for aparticular disorder. The dosage regimes necessary for prophylacticversus therapeutic treatment can vary, and will need to be designed forthe specific use and disorder treated.

In some methods, two or more agents (e.g., monoclonal antibodies withdifferent binding specificities) are administered simultaneously, inwhich case the dosage of each agent administered falls within the rangesindicated. Intervals can also be irregular as indicated by measuringreceptor saturation levels or by following other indicia of the diseaseprocess.

Those of skill will readily appreciate that dose levels can vary as afunction of the specific agent, the severity of the symptoms and thesusceptibility of the subject to side effects. Some of the specificagents are more potent than others. Preferred dosages for a given agentare readily determinable by those of skill in the art by a variety ofmeans. A preferred means is to measure the physiological potency of agiven agent.

In prophylactic applications, pharmaceutical compositions arechronically administered to a patient susceptible to, or otherwise atrisk of, a particular disease in an amount sufficient to eliminate orreduce the risk or delay the outset of the disease. Such an amount isdefined to be a prophylactically effective dose.

The anti alpha-4 agents of the invention can be used with effectiveamounts of other therapeutic agents against acute and chronicinflammation. Such agents include other antagonists of adhesionmolecules (e.g., other integrins, selectins, and immunoglobulin (Ig)superfamily members (see Springer, Nature (1990) 346:425-433; Osborn(1990) Cell 62:3; Hynes (1992) Cell 9:11)). Integrins are heterodimerictransmembrane glycoproteins consisting of an a chain (120-180 kDa) and aβ chain (90-110 kDa), generally having short cytoplasmic domains. Forexample, three important integrins, LFA-1, Mac-1 and P150,95, havedifferent alpha subunits, designated CD11a, CD11b and CD11c, and acommon beta subunit designated CD18. LFA-1 (α_(L) _(β) ₂) is expressedon lymphocytes, granulocyte and monocytes, and binds predominantly to anIg-family member counter-receptor termed ICAM-1 and related ligands.ICAM-1 is expressed on many cells, including leukocytes and endothelialcells, and is up-regulated on vascular endothelium by cytokines such asTNF and IL-1. Mac-1 (α_(M)β₂) is distributed on neutrophils andmonocytes, and also binds to ICAM-1. The third β2 integrin, P150,95(α_(X)β₂), is also found on neutrophils and monocytes. The selectinsconsist of L-selectin, E-selectin and P-selectin.

Other antiinflammatory agents that can be used in combination with theanti alpha-4 agents include antibodies and other antagonists ofcytokines, such as interleukins IL- 1 through IL-13, tumor necrosisfactors α and β (TNF-α and TNF-β), interferons α, β and γ, tumor growthfactor Beta (TGF-β), colony stimulating factor (CSF) and granulocytemonocyte colony stimulating factor (GM-CSF). Other antiinflammatoryagents include antibodies and other antagonists of chemokines such asMCP-1, MIP-1a, MIP-1β, RANTES, exotaxin and IL-8. Otheranti-inflammatory agents include NSAIDS, steroids and other smallmolecule inhibitors of inflammation. Formulations, routes ofadministration and effective concentrations of agents for combinedtherapies are as described above for the humanized antibodies againstalpha-4 integrin.

Treatment Regimes

The invention further relates to regimes for the treatment of rheumatoidarthritis. These regimes may include administering to a subject about 2mg to about 20 mg of methotrexatexate as well as an antibody to alpha-4integrin or an immunologically active antigen binding fragment thereof.Alternatively, the regimen may include administering to a subject about2 mg to about 20 mg of methotrexatexate as well as a small moleculealpha-4 integrin antagonist. Preferably, the subject is a mammal. Morepreferably, the mammal is human. Preferably, the antibody to alpha-4integrin or immunologically active antigen binding fragment isadministered in an amount of about 0.01 mg/kg of body weight to about 50mg/kg of body weight. Preferably, the small molecule alpha-4 integrinantagonist is administered in an amount of about 2 mg to about about 20mg of methotrexate and about 0.01 mg/kg of body weight to about 100mg/kg of body weight of a compound.

The regimen may be administered at any time of day or week, and in anyorder, such that a therapeutically effective does is achieved. Theantibodies, fragments or compounds may be administered prior to,followings or concurrently with the methotrexate such that the benefitsdof the combination therapy are achieved. The amount of methotrexateadministered per week should not exceed 20 mg.

EXAMPLES

The following examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the present invention, and is not intended to limit thescope of what the inventors regard as their invention nor is it intendedto represent that the experiments below are all or the only experimentsperformed. Efforts have been made to ensure accuracy with respect tonumbers used (e.g., amounts, temperature, etc.) but some experimentalerrors and deviations should be accounted for. Unless indicatedotherwise, parts are parts by weight, molecular weight is weight averagemolecular weight, temperature is in degrees Centigrade, and pressure isat or near atmospheric.

Synthesis of Compounds

Synthesis of Compounds of Formulae I and II

In the examples below, if an abbreviation is not defined above, it hasits generally accepted meaning. Further, all temperatures are in degreesCelsius (unless otherwise indicated). The following Methods were used toprepare the compounds set forth below as indicated.

Method 1 N-Tosylation Procedure

N-Tosylation of the appropriate amino acid was conducted via the methodof Cupps, Boutin and Rapoport J. Org. Chem. 1985, 50, 3972.

Method 2 Methyl Ester Preparation Procedure

Amino acid methyl esters were prepared using the method of Brenner andHuber Helv. Chim. Acta 1953, 36, 1109.

Method 3 BOP Coupling Procedure

The desired dipeptide ester was prepared by the reaction of a suitableN-protected amino acid (1 equivalent) with the appropriate amino acidester or amino acid ester hydrochloride (1 equivalent),benzotriazol-1-yloxy-tris(dimethylamino)phosphonium hexafluorophosphate[BOP] (2.0 equivalent), triethylamine (1.1 equivalent), and DMF. Thereaction mixture was stirred at room temperature overnight. The crudeproduct is purified flash chromatography to afford the dipeptide ester.

Method 4 Hydrogenation Procedure I

Hydrogenation was performed using 10% palladium on carbon (10% byweight) in methanol at 30 psi overnight. The mixture was filteredthrough a pad of Celite and the filtrate concentrated to yield thedesired amino compound.

Method 5 Hydrolysis Procedure I

To a chilled (0° C.) THF/H₂O solution (2:1, 5-10 mL) of the appropriateester was added LiOH (or NaOH) (0.95 equivalents). The temperature wasmaintained at 0C and the reaction was complete in 1-3 hours. Thereaction mixture was extracted with ethyl acetate and the aqueous phasewas lyophilized resulting in the desired carboxylate salt.

Method 6 Ester Hydrolysis Procedure II

To a chilled (0° C.) THF/H₂O solution (2:1, 5-10 mL) of the appropriateester was added LiOH (1.1 equivalents). The temperature was maintainedat 0° C. and the reaction was complete in 1-3 hours. The reactionmixture was concentrated and the residue was taken up into H₂O and thepH adjusted to 2-3 with aqueous HCl. The product was extracted withethyl acetate and the combined organic phase was washed with brine,dried over MgSO₄, filtered and concentrated to yield the desired acid.

Method 7 Ester Hydrolysis Procedure III

The appropriate ester was dissolved in dioxane/H₂O (1:1) and 0.9equivalents of 0.5 N NaOH was added. The reaction was stirred for 3-16hours and than concentrated. The resulting residue was dissolved in H₂Oand extracted with ethyl acetate. The aqueous phase was lyophilized toyield the desired carboxylate sodium salt.

Method 8 Sulfonylation Procedure I

To the appropriately protected aminophenylalanine analog (11.2 mmol),dissolved in methylene chloride (25 ml) and cooled to −78° C. was addedthe desired sulfonyl chloride (12 mmol) followed by dropwise addition ofpyridine (2 mL). The solution was allowed to warm to room temperatureand was stirred for 48 hr. The reaction solution was transferred to a250 mL separatory funnel with methylene chloride (100 mL) and extractedwith 1N HCl (50 mL×3), brine (50 mL), and water (100 mL). The organicphase was dried (MgSO₄) and the solvent concentrated to yield thedesired product.

Method 9 Reductive Amination Procedure

Reductive amination of Tos-Pro-p-NH2-Phe with the appropriate aldehydewas conducted using acetic acid, sodium triacetoxyborohydride, methylenechloride and the combined mixture was stirred at room temperatureovernight. The crude product was purified by flash chromatography.

Method 10 BOC Removal Procedure

Anhydrous hydrochloride (HCI) gas was bubbled through a methanolicsolution of the appropriate Boc-amino acid ester at 0° C. for 15 minutesand the reaction mixture was stirred for three hours. The solution wasconcentrated to a syrup and dissolved in Et₂O and reconcentrated. Thisprocedure was repeated and the resulting solid was placed under highvacuum overnight.

Method 11 tert-Butyl Ester Hydrolysis Procedure-I

The tert-butyl ester was dissolved in CH₂Cl₂ and treated with TFA. Thereaction was complete in 1-3 hr at which time the reaction mixture wasconcentrated and the residue dissolved in H₂O and lyophilized to yieldthe desired acid.

Method 12 EDC Coupling Procedure I

To a CH₂Cl₂ solution (5-20 mL) of N-(toluene-4-sulfonyl)-L-proline (1equivalent), the appropriate amino acid ester hydrochloride (1equivalent), N-methylmorpholine (1. 1-2.2 equivalents) and1-hydroxybenzotriazole (2 equivalents) were mixed, placed in an ice bathand 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide (1.1 equivalents)added. The reaction was allowed to rise to room temperature and stirredovernight. The reaction mixture was poured into H₂O and the organicphase was washed with sat. NaHCO₃, brine, dried (MgSO₄ or Na₂SO₄),filtered and concentrated. The crude product was purified by columnchromatography.

Method 13 EDC Coupling Procedure II

To a DMF solution (5-20 mL) of the appropriate N-protected amino acid (1equivalent), the appropriated amino acid ester hydrochloride (1equivalent), Et₃N (1.1 equivalents) and 1-hydroxybenzotriazole (2equivalents) were mixed, placed in an ice bath and1-(3-dimethylaminopropyl)-3-ethyl carbodiimide (1.1 equivalents) added.The reaction was allowed to rise to room temperature and stirredovernight. The reaction mixture was partitioned between EtOAc and H₂Oand the organic phase washed with 0.2 N citric acid, H₂O, sat. NaHCO₃,brine, dried (MgSO₄ or Na₂SO₄), filtered and concentrated. The crudeproduct was purified by column chromatography or preparative TLC.

Method 14 Sulfonylation Procedure II

The appropriate sulfonyl chloride was dissolved in CH₂Cl₂ and placed inan ice bath. L-Pro-L-Phe-OMe.HCl (1 equivalent) and Et₃N (1.1equivalent) was added and the reaction allowed to warm to roomtemperature and stirred overnight under an atmosphere of nitrogen. Thereaction mixture was concentrated and the residue partitioned betweenEtOAc and H₂O and the organic phase washed with sat. NaHCO₃, brine,dried (MgSO₄ or Na₂SO₄), filtered and concentrated. The crude productwas purified by column chromatography or preparative TLC.

Method 15 Sulfonylation Procedure III

To a solution of L-Pro-L-4-(3-dimethylaminopropyloxy)-Phe-OMe [preparedusing the procedure described in Method 10] (1 equivalent) in CH₂Cl₂ wasadded Et₃N (5 equivalents) followed by the appropriate sulfonyl chloride(1 equivalent). The reaction was allowed to warm to room temperature andstirred overnite under an atmosphere of nitrogen. The mixture wasconcentrated, dissolved in EtOAc, washed with sat. NaHCO₃ and 0.2 Ncitric acid. The aqueous phase was made basic with solid NaHCO₃ and theproduct extracted with EtOAc. The organic phase was washed with brine,dried (MgSO₄ or Na₂SO₄), filtered and concentrated. The crude methylester was purified by preparative TLC. The corresponding acid wasprepared using the procedure described in Method 7.

Method 16 Hydrogenation Procedure II

To a methanol (10-15 mL) solution of the azlactone was added NaOAc (1equivalent) and 10% Pd/C. This mixture was placed on the hydrogenator at40 psi H₂. After 8-16 hours, the reaction mixture was filtered through apad of Celite and the filtrate concentrated to yield thedehydrodipeptide methyl ester. The ester was dissolved in dioxane/H₂O(5-10 mL), to which was added 0.5 N NaOH (1.05 equivalents). Afterstirring for 1-3 hours, the reaction mix was concentrated and theresidue was redissolved in H₂O and washed with EtOAc. The aqueous phasewas made acidic with 0.2 N HCl and the product was extracted with EtOAc.The combined organic phase was washed with brine (1×5 mL), dried (MgSO₄or Na₂SO₄), filtered and concentrated to yield the acid as approximatelya 1 :1 mixture of diastereomers.

Method 17 tert-Butyl Ester Hydrolysis Procedure I

The tert-butyl ester was dissolved in CH₂Cl₂ (5 mL) and treated with TFA(5 mL). The reaction was complete in 1-3 hours at which time thereaction mixture was concentrated and the residue dissolved in H₂O andconcentrated. The residue was redissolved in H₂O and lyophilized toyield the desired product.

Example 1 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanineEthyl Ester

The title compound was prepared following the procedure outlined for thepreparation of Example 4 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR ((CD₃)₂SO): δ=8.33 (d, 1H), 7.70 (d, 2H), 7.41 (d, 2H), 7.24 (d,2H), 7.00 (d, 2H), 4.52-4.44 (m, 1H), 4.09-4.00 (m, 3H), 3.53 (bs, 2H),3.38-3.31 (m, 3H), 3.11-3.01 (m, 3H), 2.39 (s, 3H), 2.32 (bs, 4H), 2.19(s, 3H), 1.61-1.50 (m, 3H), 1.43-1.38 (m, 1H), 1.13 (t, 3H).

¹³C NMR ((CD₃)₂SO): δ=171.1, 171.1, 153.9, 149.8, 143.6, 134.1, 133.9,130.0, 129.8, 127.4, 121.5, 61.2, 60.7, 54.2, 54.1, 53.3, 49.0, 45.7,44.0, 43.4, 35.8, 30.5, 23.8, 21.0, 14.0.

Example 2 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineEthyl Ester

Into a reaction vial were combined 7.00 g (15.2 mmol, 1.0 eq)Ts-Pro-Tyr(H)-OEt and 1.86 g (15.2 mmol, 1.0 eq) DMAP. Methylenechloride (50 mL), triethylamine (2.12 mL-1.54 g, 15.2 mmol, 1.0 eq), anddimethylcarbamyl chloride (1.68 mL-1.96 g, 18.2 mmol, 1.2 eq) were thenadded. The vial was capped tightly, and the reaction solution swirled toobtain a homogeneous solution. The reaction solution was then heated to40° C. After 48 h, TLC of the resulting colorless solution indicatedcomplete conversion. The workup of the reaction solution was as follows:add 50 mL EtOAc and 50 mL hexanes to the reaction mixture, and wash with3×50 mL 0.5 mL hexanes to the reaction mixture, and wash with 3×50 mL0.5 M citric acid, 2×50 mL water, 2×50 mL 10% K₂CO₃, and 1×50 mL sat.NaCl. Dry with MgSO₄. Filter. Evaporate to obtain 8.00 g (99%) of thetitle compound as a clear oil, which solidifies upon standing.Recrystallize from 5:3:2 heptane/EtOAc/CH₂CI₂.

NMR data was as follows:

¹H NMR ((CD₃)₂SO): δ=8.32 (d, 1H), 7.70 (d, 2H), 7.41 (d, 2H), 7.23 (d,2H), 7.00 (d, 2H), 4.52-4.44 (m, 1H), 4.09-4.02 (m, 3H), 3.37-3.31 (m,1H), 3.11-2.96 (m, 3H), 3.00 (s, 3H), 2.87 (s, 3H), 2.39 (s, 3H),1.61-1.50 (m, 3H), 1.43-1.38 (m, 1H), 1.13 (t, 3H).

¹³C NMR (CD₃)₂SO): δ=171.1, 171.1, 154.0, 150.0, 143.6, 133.9, 133.9,130.0, 129.8, 127.4, 121.5, 61.2, 60.6, 53.3, 49.0, 36.3, 36.1, 35.8,30.5, 23.8, 21.0, 14.0.

Example 3 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure outlined for thepreparation of Example 4 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.72 (d, 2H), 7.36 (d, 1H), 7.33 (d, 2H), 7.16 (d,2H), 7.03 (d, 2H), 5.07 (September, 1H), 4.78 (dt, 1H), 4.08-4.05 (m,1H), 3.67 (bs, 2H), 3.57 (bs, 2H), 3.41-3.35 (m, 1H), 3.24 (dd, 1H),3.15-3.07 (m, 1H), 3.04 (dd, 1H), 3.46-2.43 (m, 7H), 2.34 (s, 3H),2.05-2.02 (m, 1H).

¹³C NMR (CDCl₃): δ=170.9, 170.4, 153.6, 150.5, 144.3, 133.2, 133.1,130.2, 130.0, 127.9, 121.7, 69.5, 62.2, 54.7, 53.4, 49.6, 46.1, 44.3,43.7, 37.2, 29.7, 24.1, 21.6, 21.6, 21.4.

Example 4 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalaninetert-Butyl Ester

Combine 41.2 g (84.34 mmol, 1.0 eq) Ts-Pro-Tyr(H)-OtBu and 17.0 g (84.34mmol, 1.0 eq) 4-nitrophenyl chloroformate. Add 700 mL CH₂Cl₂. Cap with aseptum. Attach a N₂ line. Immerse the flask in a 4:1 water/EtOH +dry iceslurry, and stir to cool to −15° C. Add 29.38 mL (21.33 g, 210.81 mmol,2.5 eq) Et₃N over five minutes-with stirring. Stir at −10 to 15° C. for1 h. Add 9.35 mL (8.45 g, 84.34 mmol, 1.0 eq) N-methyl piperazine over 3minutes with stirring. Stir overnight while warming to room temperature.Dilute with 700 mL hexanes. Wash repeatedly with 10% K₂CO₃, until noyellow color (4-nitrophenol) is seen in the aqueous layer. Wash withsat. NaCl. Dry over anhydrous MgSO₄. Filter. Evaporate. Dissolve in 500mL EtOH, and evaporate, to remove Et₃N. Repeat once. Dissolve in 400 mLEtOH, and add 600 mL water with stirring, to precipitate a solid or oil.If an oil, stir vigorously to solidify. Isolate the solid by filtration.Repeat dissolution, precipitation, and filtration, once. Rinse withwater to remove traces of yellow color. High vacuum to constant massyields the title compound as a white solid.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.72 ( d, 2H), 7.33 (d, 3H), 7.17 (d, 2H), 7.02 (d,2H), 4.71 (q, 1H), 4.09-4.06 (m, 1H), 3.67 (bs, 2H), 3.57 (bs, 2H),3.41-3.34 (m, 1H), 3.22 (dd, 1H), 3.16-3.09 (m, 1H), 3.03 (dd, 1H),2.46-2.43 (m, 7H), 2.34 (s, 3H), 2.05-2.02 (m, 1H), 1.57-1.43 (m, 3H),1.47 (s, 9H).

³C NMR (CDCl₃): δ=171.8, 169.9, 153.6, 150.4, 144.3,133.4, 133.1, 130.3,130.0, 127.9, 121.6, 82.6, 62.3, 54.5, 53.8, 49.6, 46.1, 44.3, 43.7,37.3, 29.7, 27.8, 24.1, 21.4.

Example 5 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine

The title compound was prepared from the product of Example 1 using theprocedure described in Method 7.

NMR data was as follows:

¹H NMR (CD₃OD): δ=7.74 (d, 2H), 7.42 (d, 2H), 7.26 (d, 2H), 7.04 (d,2H), 4.58-4.54 (m, 1H), 4.16-4.12 (m, 1H), 3.70 (bs, 2H) 3.53 (bs, 2H),3.43-3.31 (m, 1H), 3.26-3.13 (m, 7H), 2.82 (s, 3H), 2.43 (s, 3H),1.98-1.94 (m, 1H), 1.76-1.51 (m, 3H).

¹³C NMR (CD₃OD): δ=175.7, 173.6, 154.8, 151.6, 146.1, 136.3, 134.8,131.9, 131.3, 129.1, 122.7, 63.6, 55.9, 53.9, 50.7, 43.5, 37.6, 31.3,25.5, 21.5.

Example 6 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy-)phenylalaninen-Butyl Ester

The title compound was prepared following the procedure outlined for thepreparation of Example 2 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CD₃)₂SO: δ=8.31 (d, 1H), 7.70 (d, 2H), 7.41 (d, 2H), 7.23 (d,2H), 6.99 (d, 2H), 4.53-4.46 (m, 1H), 4.10-4.01 (m, 1H), 3.63-3.30 (m,1H), 3.10-2.96 (m, 3H), 3.00 (s, 3H), 2.88 (s, 3H), 2.39 (s, 3H),1.59-1.30 (m, 6H), 1.33-1.20 (m, 2H), 0.85 (t, 3H).

¹³CNMR(CD₃)₂SO: δ=171.4, 171.3, 154.2, 150.2, 143.7, 134.0, 130.1,130.0, 127.6, 121.7, 64.3, 61.2, 59.2, 53.4, 49.0, 36.2, 36.0, 35.8,30.0, 23.8, 21.0, 18.5, 13.5.

Example 7 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineCyclopentyl Ester

The title compound was prepared following the procedure outlined for thepreparation of Example 2 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CD₃)₂SO: δ=8.27 (d, 1H), 7.70 (d, 2H), 7.41 (d, 2H), 7.22 (d,2H), 6.99 (d, 2H), 5.04 (bs, 1H), 4.48-4.40 (m, 1H), 4.08-4.05 (m, 1H),3.34-3.30 (m, 1H), 3.09-2.95 (m, 3H), 3.00 (s, 3H), 2.88 (s, 3H), 2.39(s, 3H), 1.76-1.74 (m, 2H), 1.57-1.40 (m, 10H).

¹³C NMR (CD₃)₂SO: δ=171.3, 171.0, 154.2, 150.2, 432.7, 134.1, 130.1,130.0, 127.6, 121.6, 77.4, 61.2, 53.4, 49.0, 36.2, 36.1, 35.7, 32.0,30.5, 23.8, 23.2, 21.0.

Example 8 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure outlined for thepreparation of Example 2 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CD₃)₂SO: δ=8.18 (d, 1H), 7.71 (d, 2H), 7.41 (d, 2H), 7.23 (d,2H), 6.99 (d, 2H), 4.42 4.38 (m, 1H), 4.10-4.07 (m, 1H), 3.37-3.30 (m,1H), 3.09-2.95 (m, 3H), 3.00 (s, 3H), 2.88 (s, 3H), 2.39 (s, 3H),1.58-1.50 (m, 3H), 1.40-1.30 (m, 1H), 1.36 (s, 9H).

¹³C NMR (CD₃)₂SO: δ=171.1, 170.3, 154.2, 150.2, 143.8, 134.2, 134.1,130.2, 130.0, 127.6, 121.6, 81.0, 61.3, 53.8, 49.0, 36.3, 36.0, 35.9,30.5, 27.5, 23.8, 21.0.

Example 9 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 2 using theprocedure described in Method 7.

NMR data was as follows:

¹H NMR (CD₃)₂SO: δ=8.13 (d, 1H), 7.70 (d, 2H), 7.41 (d, 2H), 7.23 (d,2H), 6.99 (d, 2H), 4.51-4.44 (m, 1H), 4.11-4.09 (m, 1H), 3.40-3.34 (m,2H), 3.11-2.94 (m, 3H), 3.00 (s, 3H), 2.87 (s, 3H), 2.39 (s, 3H),1.59-1.36 (m, 4H).

¹³C NMR (CD₃)₂SO: δ=172.7, 171.2, 153.6, 150.2, 143.8, 134.3, 134.0,130.2, 130.0, 127.6, 121.6, 61.3, 53.2, 49.0, 36.3, 36.1, 35.9, 30.4,23.8, 21.0.

Example 10 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-3-(N,N-dimethylcarbamyloxy)phenylalanineEthyl Ester

The title compound was prepared following the procedure outlined for thepreparation of Example 2 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.74 (m, 2H), 7.70-7.36 (m, 4H), 7.24-7.14 (m, 3H),6.93-4.90 (m, 1H), 4.78-4.27 (m, 3H), 4.05-3.55 (m, .5H), 3.48-3.43 (m,.5H), 3.37-3.30 (m, 3H), 3.02-3.08 (bs, 3H), 2.99 (bs, 3H), 2.45 (s,1.5H), 2.43 (s, 1.5H), 2.12 (m, 1H), 198, 1.80 (m, 0.5M),1.62-1.44 (m,2.5H), 1.29 (t, 1.5H), 1.24 (t, 1.5H).

¹³C NMR (CDCl₃): δ=171.1, 171.0, 170.9, 154.9, 154.8, 151.8, 151.6,144.4, 144.3, 137.6, 137.1, 133.1, 132.9, 130.0, 129.9, 129.5, 129.2,127.9, 127.9, 126.5, 126.1, 122.9, 122.7, 120.7, 120.5.

Example 11 Synthesis ofN-(Toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure outlined for thepreparation of Example 2 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.76 (d, 2H), 7.35 (d, 2H), 7.22 (d, 2H), 7.01 (m,3H), 5.05 (m, 1H), 4.85 (m, 1H), 4.57 (d, 1H), 4.38 (d, 1H), 3.86 (s,1H), 3.19-3.00 (m, 2H), 3.09 (s, 3H), 3.01 (s, 3H), 2.45 (s, 3H), 1.24(t, 6H), 1.16 (s, 3H), 1.09 (s, 3H).

¹³C NMR (CDCl₃): δ=170.3, 168.4, 154.9, 150.6, 144.8, 132.9, 132.8,130.3, 130.0, 128.2, 121.7, 73.4, 69.5, 54.5, 53.2, 50.4, 37.7, 36.5,36.3, 29.0, 23.8, 21.5, 21.4.

Example 12 Synthesis ofN-(Toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure outlined for thepreparation of Example 2 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.75 (d, 2H), 7.34 (d, 2H), 7.23 (d, 2H), 7.05-6.98(m, 3H), 4.76 (m, 1H), 4.56 (d, 1H), 4.40 (d, 1H), 3.85 (s, 1H),3.09-3.00 (m, 8H), 2.44 (s, 3H), 1.43 (s, 3H), 1.16 (s, 3H), 1.09 (s,3H).

¹³C NMR (CDCl₃): δ=169.8, 168.3, 154.9, 150.6, 144.8, 133.2, 132.9,130.4, 130.0, 128.2, 121.6, 82.6, 73.4, 54.6, 53.8, 50.4, 37.8, 36.5,36.3, 29.0, 27.7, 23.8, 21.5.

Example 13 Synthesis ofN-(Toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 11 using theprocedure described in Method 7.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.76 (d, 2H), 7.35 (d, 2H), 7.25 (d, 2H), 7.14 (d,1H), 7.02 (d, 2H), 5.17 (br s, 1H), 4.89 (m, 1H), 4.56 (d, 1H), 4.40 (d,1H), 3.90 (s, 1H), 3.30-3.00 (m, 8H), 2.43 (s, 3H), 1.09 (s, 6H).

¹³C NMR (CDCl₃): δ=172.7, 169.3, 155.2, 150.6, 144.9, 133.1, 132.7,130.5, 130.1, 128.1, 121.9, 73.3, 54.5, 53.3, 50.5, 36.9, 36.6, 36.4,29.0, 23.7, 21.5.

Example 14 Synthesis ofN-(Toluene-4-sulfonyl)-L-[(1,1-dioxo)thiamorpholin-3-carbonyl]-L4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

L-Thiamorpholine-5-carboxylic acid was prepared by the method of Larssonand Carlson (Acta Chemica Scan. 1994, 48, 517-525).N-(Toluene-4-sulfonyl)-L-thiamorpholine-5-carboxylic acid was preparedusing the procedure described in Method 1 and was then coupled tot-butyl tyrosine in DMF in the presence of BOP and NMM, to give afteraqueous workup and flash chromatographyN-(Toluene-4-sulfonyl)-L-[thiamorpholin-3-carbonyl]-L-4-phenylalaninetert-butyl ester.

Formation of the 4-(N,N-dimethylcarbamyloxy) group was per Example 2above and oxidation of the thiamorpholino group to the1,1-dioxo-thiamorpholino group was per Larsson and Carlson (Acta ChemicaScan. 1994, 48, 522).

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.68 (d, 2H), 7.37 (d, 2H), 7.08 (m, 4H), 6.73 (d,1H), 5.11 (m, 1 H), 4.62 (m, 1H), 4.23 (m, 1H), 4.00 (m, 1H), 3.82 (m,1H), 3.14 (s, 3H), 3.03 (s, 3H), 2.80 (m, 5H), 2.44 (s, 3H), 1.48 (s,9H).

¹³C NMR (CDCl₃): δ=171.3, 169.9, 164.4, 145.6, 135.4, 132.6, 130.8,130.4, 127.3, 121.9, 83.0, 56.1, 53.8, 49.4, 48.7, 44.5, 42.0, 36.9,36.6, 36.4, 27.8, 21.5.

Example 15 Synthesis ofN-(Toluene-4-sulfonyl)-L-[(1,1-dioxo)thiamorpholin-3-carbonyl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 14 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ=7.77 (d, 2H), 7.40 (d, 2H), 7.22 (d, 2H), 7.00 (d,2H), 5.19 (m, 1H), 4.65 (m, 1H), 4.30 (m, 1H), 3.95 (m, 1H), 3.61 (m,1H), 3.20 (m, 5H), 3.09 (s, 3H), 2.97 (s, 3H), 2.43 (s, 3H).

¹³C NMR (CD₃OD): δ=174.1, 168.0, 157.0, 152.0, 146.4, 137.7, 135.3,131.7, 131.6, 128.8, 123.0, 57.1, 54.8, 51.1, 50.9, 48.0, 47.7, 43.2,37.4, 36.8, 36.7, 21.5.

Example 16 Synthesis ofN-(Toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure outlined for thepreparation of Example 4 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.74 (d, 2H), 7.33 (d, 2H), 7.25 (d, 2H), 7.20-7.00(m, 3H), 4.74 (m, 1H), 4.55 (d, 1H), 4.38 (d, 1H), 3.83 (s, 1H), 3.66(br m, 2H), 3.57 (br m, 2H), 3.08-3.05 (m, 2H), 2.45-2.42 (m, 7H), 2.33(s, 3H), 1.42 (s, 9H), 1.15 (s, 3H), 1.08 (s, 3H).

¹³C NMR (CDCl₃): δ=169.7, 168.2, 153.6, 150.3, 144.7, 133.3, 132.7,130.4, 129.9, 128.1, 121.5, 82.6, 73.4, 54.5, 53.7, 50.4, 46.0, 44.2,43.6, 37.7, 28.9, 27.7, 23.8, 21.4.

Example 17 Synthesis ofN-(Toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine

The title compound was prepared from the product Example 16 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CDCl₃): δ=8.31 (d, 1H), 7.72 (d, 2H), 7.42-7.35 (m, 4H), 7.08(d, 2H), 4.90-4.68 (m, 1H), 4.64-4.61 (m, 1H), 4.47-4.44 (m, 1H), 4.01(s, 1H), 3.36-3.32 (br m, 4H), 3.27-3.25 (m, 1H), 3.22-3.10 (m, 1H),2.94 (s, 3H), 2.43 (s, 3H), 1.14 (s, 3H), 1.07 (s, 3H).

Example 18 Synthesis ofN-(Toluene-4-sulfonyl)sarcosyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure outlined for thepreparation of Example 2 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.66 (d, 2H), 7.34 (d, 2H), 7.18 (d, 2H), 7.07 (d,2H), 6.98 (d, 1H), 5.03 (m, 1H), 4.81 (m, 1H), 3.69 (d, 1H), 3.49 (d,1H), 3.08 (m, 2H), 3.04 (s, 3H), 2.99 (s, 3H), 2.63 (s, 3H), 2.43 (s,3H).

¹³C NMR (CDCl₃): δ=167.4, 154.9, 150.8, 144.4, 132.6, 130.2, 130.1,127.7, 122.0, 110.9, 69.5, 57.3, 53.9, 53.0, 37.1, 36.6, 21.6, 21.4.

Example 19 Synthesis ofN-(Toluene4-sulfonyl)sarcosyl-L4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure for thepreparation of Example 2 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.67 (d, 2H), 7.34 (d, 2H), 7.19 (d, 2H), 7.03 (d,2H), 6.98 (d, 1H), 4.76 (m, 1H), 3.67 (q, 1H), 3.06 (m, 2H), 3.16 (s,3H), 2.99 (s, 3H), 2.64 (s, 3H), 2.43 (s, 3H), 1.42 (s, 9H).

¹³C NMR (CDCl₃): δ=170.0, 137.2, 154.9, 150.7, 144.3, 133.2, 132.9,130.3, 130.0, 127.7, 121.9, 82.6, 83.9, 53.3, 37.2, 36.6, 36.4, 27.9,21.4.

Example 20 Synthesis ofN-(Toluene-4-sulfonyl)sarcosyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 18 using theprocedure described in Method 7.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.41 (d, 2H), 7.10 (d, 2H), 6.98 (d, 2H), 6.75 (d,2H), 4.42 (m, 1H), 3.43 (m, 2H), 3.04 (m, 2H), 2.80 (s, 3H), 2.69 (s,3H), 2.33 (s, 3H), 2.14 (s, 3H).

¹³C NMR (CDCl₃): δ=174.2, 170.2, 156.9, 151.9, 145.6, 135.5, 135.2,131.4, 131.1, 128.9, 123.0, 54.6, 54.0, 37.4, 36.8, 36.7, 21.4.

Example 21 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylaminosulfonyloxy)phenylalaninetert-Butyl Ester

Substituting dimethysulfamoyl chloride for dimethylcarbamyl chloride,and following the method for the preparation of Example 2, gave thetitle compound.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.72 (d, 2H), 7.34 (d, 2H), 7.21 (s, 4H), 4.69 (m,1H), 4.04 (m, 1H), 3.4 (m, 1H), 3.24 (m, 3H), 2.96 (s, 6H), 2.42 (s,3H), 2.02 (m, 1H), 1.45 (m, 13H).

¹³C NMR (CDCl₃): δ=166.3, 165.3, 144.8, 140.0, 130.9, 126.4, 125.6,123.5, 117.3, 95.5, 78.3, 57.8, 49.2, 45.2, 34.2, 32.9, 25.0, 23.4,19.7, 17.1.

Example 22 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylaminosulfonyloxy)phenylalanine

The title compound was prepared from the product of Example 21 using theprocedure described in method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ=7.73 (d, 2H), 7.41 (d, 2H), 7.38 (d, 2H), 7.22 (d,2H), 4.69 (m, 1H), 4.11 (m, 1H), 3.41 (m, 2H), 3.19 (m, 2H), 2.94 (s,6H), 2.41 (s, 3A), 1.78 (m, 1H), 1.61 (m, 3H).

¹³C NMR (CD₃OD): δ=174.3, 174.0, 150.8, 145.9, 137.3, 135.1, 132.1,131.2, 129.1, 123.1, 63.3, 54.6, 50.6, 39.1, 37.5, 31.6, 25.3, 21.5.

Example 23 Synthesis ofN-(Toluene-4-sulfonyl)-sarcosyl-L-(4-morpholinecarbamyloxy)pheny!alanine t-butyl ester

Substituting sacrosine for L-proline in the preparation ofTs-Pro-Tyr(H)-O-t-butyl ester and substitution of 4-morpholinecarbonylchloride for dimethylcarbamyl chloride, and following the method for thepreparation of Example 2, gave the title compound as a white solid.

NMR data was as follows:

¹NMR (CDCl₃): δ7.61 (d, 2H), 7.28 (d, 2H), 7.16 (d, 2H), 7.02 (d, 2H),4.69 (m, 1H), 3.67 (m, 8H), 3.58 (m, 1H), 3.48 (m, 1H), 3.06 (m, 2H),2.59 (s, 3H), 2.36 (s, 3H), 1.26 (s, 9H).

¹³C NMR (CDCl₃): δ169.7, 167.1, 153.5, 150.1, 144.1, 133.1, 133.0,133.0, 130.1, 129.8, 127.4, 121.6, 82.6, 66.3, 53.6, 53.1, 44.5, 43.7,36.9, 36.4, 27.6, 21.2.

Example 24 Synthesis ofN-(Toluene4-sulfonyl)sarcosyl-L4-(isonipecotoyloxy)phenylalanine

The title compound was prepared from the product of Example 23 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ=7.30 (d, 2H), 7.02 (d, 2H), 6.88 (d, 2H), 6.67 (d,2H), 4.33 (m, 1H), 3.32 (m, 3H), 3.25 (m, 2H), 3.12 (m, 3H), 2.89 (m,1H), 2.70 (m, 3H), 2.22 (s, 3H), 2.03 (s, 3H).

¹³C NMR (CD₃OD): δ=174.2, 170.3, 155.6, 151.7, 145.6, 135.8, 135.2,131.5, 131.1, 128.9, 123.0, 67.5, 54.6, 54.0, 37.4, 36.8, 21.5.

Example 25 Synthesis ofN-(Toluene-4-sulfonyl)-L-[(1,1-dioxo)thiamorpholin-3-carbonyl]-L-4-(morpholin-4-ylcarbonyloxy)phenylalaninetert-Butyl Ester

Substitution of 4-morpholinecarbonyl chloride for dimethylcarbamylchloride, and following the methods for the preparation of Example2 and14, gave the title compound as a white solid.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.76 (d, 1H), 7.68 (d, 1H), 7.37 (m, 2H), 7.14 (m,2H), 7.05 (m, 1H), 6.97 (d, 1H), 6.80 (d, 0.5H), 6.57 (d, 0.5H), 5.09(m, 0.5H), 4.91 (m, 0.5H), 4.75 (m, 0.5H), 4.62 (m, 0.5H), 4.25 (m,0.5H), 4.09 (m, 2H), 3.79 (m, 4H), 3.65 (m, 4H), 2.91 (s, 3H), 2.44 (s,3H), 1.69 (s, 4H), 1.44 (s, 5H).

¹³C NMR (CDCl₃): δ=170.0, 169.8, 164.8, 164.4, 153.7, 150.4, 145.6,145.4, 135.4, 135.3, 132.9, 130.8, 130.7, 130.5, 130.4, 127.5, 127.2,122.1, 121.8, 83.01, 82.8, 66.4, 56.1, 56.1, 53.7, 53.6, 49.5, 49.3,48.6, 44.7, 43.9, 42.0, 41.6, 36.9, 36.3, 27.8, 21.5.

Example 26 Synthesis ofN-(Toluene-4-sulfonyl)-L-[(1,1-dioxo)thiamorpholin-3-carbonyl]-L4-(morpholin4-ylcarbonyloxy)phenylalanine

The title compound was prepared from the product of Example 25 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ=7.67 (m, 2H), 7.32 (m, 2H), 7.08 (m, 2H), 6.93 (m,2H), 5.09 (m, 1H), 4.54 (m, 1H), 4.19 (m, 0.5H), 4.02 (m, 0.5H), 3.81(m, 0.5H), 3.66 (m, 8H), 2.99 (m, 7H), 2.32 (s, 3H).

¹³C NMR (CD₃OD): δ=174.0, 168.0, 155.7, 151.9, 151.8, 146.6, 146.4,137.5, 135.5, 135.3, 131.7, 131.6, 131.6, 128.8, 123.3, 122.9, 67.6,57.3, 57.1, 54.8, 51.1, 50.9, 50.6, 46.0, 45.3, 45.2, 43.0, 37.4, 37.0,21.5.

Example 27 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure outlined for thepreparation of Example 2 and substitution of appropriate startingmaterials.

NMR data was as follows:

³C NMR (CDCl₃): δ=7.87-7.83 (m, 2H), 7.26-7.13 (m, 5H), 4.74-4.69 (m,1H), 4.05 (m, 1H), 3.36 (m, 1H), 3.24-3.17 (m, 1H), 3.11-3.01 (m, 4H),2.97 (s, 3H), 2.05-2.02 (m, 1H), 1.60-1.47 (m, 3H), 1.46 (s, 9H).

¹³C NMR (CDCl₃): δ=170.6, 170.0, 165.7, 154.9, 150.6, 133.2, 132.4,130.7, 130.2, 121.7, 116.7, 82.7, 62.3, 53.7, 49.6, 37.2, 36.6, 36.4,29.9, 27.9, 24.2.

Example 28 Synthesis ofN-(Toluene-4-sulfonyl)sarcosyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure outlined for thepreparation of Example 4 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=8.17 (d, 1H), 7.59 (d, 2H), 7.26 (d, 2H), 7.13 (d,2H), 7.00 (d, 2H), 4.66 (m, 1H), 3.60 (m, 6H), 3.04 (m, 2H), 2.56 (s,3H), 2.40 (m, 7H), 2.34 (s, 3H), 1.41 (s, 9H).

¹³C NMR (CDCl₃): δ=169.7, 167.0, 153.4, 150.2, 144.0, 133.0, 132.9,130.1, 129.8, 127.4, 121.6, 82.2, 54.3, 53.5, 53.1, 45.8, 44.2, 43.5,36.9, 27.6, 21.2.

Example 29 Synthesis ofN-(Toluene-4-sulfonyl)-L-(1,1-dioxo-5,5-dimethyl)-thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The product of Example 12 was oxidized by the method of Larsson andCarlson (Acta Chemica Scan. 1994, 48, 517-525), yielding the titlecompound as a white solid.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.73 (d, 2H), 7.36 (d, 2H), 7.21 (d, 2H), 7.06-6.95(m, 3H), 4.79 (m, 1H), 4.38 (dd, 2H), 4.10 (s, 1H), 3.18-2.95 (m, 8H),2.43 (s, 3H), 1.45 (s, 9H), 1.33 (s, 3H), 1.08 (s, 3H).

¹³C NMR (CDCl₃): δ=169.8, 166.2, 154.9, 120.7, 145.8, 133.0, 131.9,130.2, 128.5, 121.9, 82.9, 68.0, 60.9, 59.3, 53.9, 37.5, 36.6, 36.3,27.7, 21.6, 19.3, 18.5.

Example 30 Synthesis ofN-(1-Methylimidazolyl-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 106 usingthe procedure described in Method 11.

NMR data was as follows:

¹H NMR (CDCl₃): δ=8.07 (d, 1H), 7.75 (s, 1H), 7.71 (s, 1H), 7.25 (d,2H), 7.01 (d, 2H), 4.71-4.66 (m, 1H), 4.28-4.24 (m, 1H), 3.77 (s, 3H),3.42-3.05 (m, 3H), 3.09 (s, 3H), 2.96 (s, 3H), 1.84-1.69 (m, 2H),1.61-1.54 (m, 2H).

¹³C NMR (CDCl₃): δ=174.4, 174.1, 156.9, 151.9, 141.8, 137.7, 135.6,131.6, 127.6, 122.9, 63.7, 54.7, 50.8, 37.4, 36.8, 36.7, 34.3, 31.6,25.4.

Preparative Example A Synthesis of2-(Saccharin-2-yl)propionoyl-L4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

N-(Benzisothiazolone)-L-alanyl-L-tyrosine t-butyl ester was prepared byfirst combining sodium hydride (washed free of mineral oil) in THFchilled to 0° C., and a solution ofN-(2-methoxycarbonyl)sulfonyl-L-alanine-L-tyrosine t-butyl ester in THFwhich was added dropwise. The reaction was stirred at 0° C. for one hourand then at room temperature for two hours. The reaction mixture wasextracted with EtOAc and 0.2 N HCl, the combined EtOAc layers werewashed successively with 0.2 N HCl, sat. NaHCO₃, and sat. NaCl. Theorganic layer was dried over MgSO₄, filtered and concentrated. Theresidue was filtered by silica gel chromatography to affordN-(benzisothiazolone)-L-alanyl-L-tyrosine t-butyl ester.

The title compound was then prepared following the procedure describedin Example 2.

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHz) (1:1 mixture of diastereomers) δ=8.15 (m, 2H);8.5 (m, 3H); 7.20 (m, 2H); 6.95 (m, 2H); 4.75 (m, 1H); 4.30 (m, 1H);3.05 (s, 3H); 2.95 (m, 2H); 2.90 (s, 3H); 1.75 and 1.65 (two d, 3H);1.30 and 1.35 (two s, 9H).

Example 31 Synthesis ofN-(Toluene-4-sulfonyl)-L-(1,1-dioxo-5,5-dimethyl)-thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 29 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.75 (m, 3H), 7.29 (m, 4H), 7.08 (d, 2H), 4.95 (m,1H), 4.46-4.20 (m, 3H), 3.17 (s, 3H), 3.30-3.10 (m, 2H), 3.02 (s, 3H),2.43 (s, 3H), 1.15 (s, 3H), 0.88 (s, 3H).

¹³C NMR (CDCl₃): δ=127.2, 167.5, 155.8, 150.3, 145.4, 133.6, 132.6,130.8, 130.2, 128.3, 121.9, 67.9, 65.8, 60.8, 53.9, 36.8, 36.6, 35.8,21.6, 18.8, 15.0.

Example 32 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-prolyl-L4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 27 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.88-7.84 (m, 2H), 7.54 (d, 1H), 7.26-7.18 (m, 4H),7.01 (d, 2H), 6.92 (s, 3H), 4.88-4.83 (m, 1H), 4.14-4.11 (m, 1H),3.39-3.29 (m, 2H), 3.13 (m, 2H), 3.00 (s, 3H), 2.99 (s, 3H), 1.92-1.89(m, 1H), 1.59-1.43 (m, 3H).

¹³C NMR (CDCl₃): δ=173.1, 172.4, 165.6, 155.5, 150.4, 133.2, 131.9,130.6, 130.3, 121.8, 116.6, 61.9, 53.1, 49.6, 36.6, 36.3, 30.2, 23.9.

Example 33 Synthesis ofN-(Toluene-4-sulfonyl)-D-prolyl-L-4-(4-methylpiperazin-1-yl)phenylalaninet-butyl Ester

The title compound was prepared following the procedure outlined for thepreparation of Example 4 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.72 (d, 2H), 7.33 (d-, 3H-), 7-.1-7 (d, 2H), 7.02 (d,2H), 4.71 (q, 1H), 4.09-4.06 (m, 1H), 3.67 (bs, 2H), 3.57 (bs, 2H),3.41-3.34 (m, 1H), 3.22 (dd, 1H), 3.16-3.09 (m, 1H), 3.03 (dd, 1H),2.46-2.43 (m, 7H), 2.05-2.02 (m, 1H), 1.57-1.43 (m, 3H), 1.47 (s, 9H).

¹³C NMR (CDCl₃): δ=170.8, 169.9, 153.6, 150.4, 144.3, 133.4, 133.1,130.3, 130.0, 127.9, 121.6, 82.6, 62.3, 54.5, 53.8, 49.6, 46.1, 44.3,43.7, 37.3, 29.7, 27.8, 24.1, 21.4.

Example 34 Synthesis ofN-(Toluene-4-sulfonyl)-N-methyl-L-alanyl-L4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalaninet-butyl ester

The title compound was prepared following the procedure outlined for thepreparation of Example 4 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.68 (d, 2H), 7.31 (d, 2H), 7.17 (d, 2H), 7.04 (d,2H), 6.86 (d, 1H), 4.65 (m, 1H), 4.47 (q, 1H), 3.71-3.53 (m, 4H),3.24-2.92 (m, 2H), 2.50-2.40 (m, 10H), 2.35 (s, 3H), 1.45 (s, 9H), 0.92(d, 3H).

¹³C NMR (CDCl₃): δ=170.1, 169.9, 153.6, 150.4, 143.9, 135.6, 133.3,130.2, 129.9, 127.2, 121.8, 82.4, 55.4, 54.6, 53.6, 46.0, 44.2, 43.7,37.2, 29.6, 27.8, 21.4, 11.5.

Example 35 Synthesis ofN-(4-Nitrobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure outlined for thepreparation of Example 2 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=8.38-8.34 (m, 2H), 8.05-8.00 (m, 2H), 7.16-2.12 (m,2H), 7.03-6.94 (m, 3H), 4.74-4.68 (m, 1H), 4.15-4.14 (m, 1H), 3.41-3.32(m, 1H), 3,23-3.14 (m, 2H), 3.08 (s, 3H), 3.03 (m, 1H), 2.98 (s, 3H),2.05 (m, 1H), 1.66-1.48 (m, 3H), 1.47 (s, 9H).

¹³C NMR (CDCl₃): δ=170.0, 169.9, 154.8, 150.6, 150.4, 142.4, 132.9,130.2, 129.0, 124.5, 121.6, 82.7, 62.2, 53.4, 49.4, 37.0, 36.5, 36.2,30.1, 27.7, 24.1.

Example 36 Synthesis ofN-(Toluene-4-sulfonyl)-L-[(1,1-dioxo)-thiamorpholin-3-carbonyl]-L-4-(N,N-dimethylaminosulfonyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared using the procedure described for thepreparation of Example 21 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.73 (d, 1H), 7.67 (d, 1H), 7.35 (m, 2H), 7.27 (m,2H), 6.88 (d, 1H), 6.66 (d, 1H), 5.08 (m, 0.5H), 4.97 (m, 0.5H), 4.71(m, 0.5H), 4.61 (m, 0.5H), 4.25 (m, 0.5H), 4.03 (m, 1H), 3.21-3.04 (m,4H), 2.89 (s, 3H), 2.83 (s, 3H), 2.78 (m, 3H), 2.42 (s, 3H), 1.44 (s,4.5H), 1.38 (s, 4.5H).

¹³C NMR (CDCl₃): δ=169.8, 169.6, 164.9, 164.5, 149.3, 149.1, 145.6,145.4, 135.4, 135.0, 134.6, 130.9, 130.6, 130.5, 127.4, 127.2, 122.0,121.8, 83.0, 83.0, 56.0, 53.7, 49.2, 49.1, 48.5, 41.9, 41.4, 38.6, 36.8,36.2, 27.7, 21.5.

Example 37 Synthesis ofN-(Toluene-4-sulfonyl)sarcosyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalaninetert-Butyl Ester

Substituting thiomorpholine for N-methylpiperazine, and following themethod for the preparation of Example 4, gave the title compound.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.65 (d, 2H), 7.33 (d, 2H), 7.20 (d, 2H), 7.04 (d,2H), 4.76 (m, 1H), 3.89 (m, 4H), 3.68 (d, 1H), 3.48 (d, 1H), 3.10 (m,2H), 2.66 (m, 7H), 2.41 (s, 3H), 1.43 (s, 9H).

¹³C NMR (CDCl₃): δ=169.9, 167.2, 153.5, 150.3, 144.3, 133.1, 130.3,130.0, 127.6, 121.8, 82.5, 53.8, 53.3, 47.0, 36.4, 37.2, 36.6, 27.8,27.3, 27.0, 21.4.

Example 38 Synthesis ofN-(Toluene-4-sulfonyl)-L-N-methylalanyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine

The title compound was prepared from the product of Example 34 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.65 (d, 2H), 7.34 (d, 2H), 7.27 (d, 2H), 7.09 (d,2H), 4.64-4.50 (m, 2H), 4.48-4.23 (m, 2H), 3.60-2.96 (m, 8H), 2.92 (s,3H), 2.55 (s, 3H), 2.40 (s, 3H), 0.93 (d, 3H).

¹³C NMR (CDCl₃): δ=174.3, 173.1, 154.9, 151.6, 145.5, 137.0, 136.1,131.6, 131.2, 128.5, 123.1, 56.4, 54.8, 54.0, 43.8, 37.3, 30.2, 21.5,13.2.

Example 39 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L4-(1,1-dioxothiomorpholin-4-ylcarbonyloxy)phenylalanine

The title compound was prepared from the product of Example 81 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ=8.03 (m, 1H), 7.73 (d, 2H), 7.41 (d, 2H), 7.28 (d,2H), 7.08 (d, 2H), 4.70-4.65 (m, 1H), 4.12-4.00 (m, 5H), 3.38-3.36 (m,1H), 3.31-3.06 (m, 7H), 2.43 (s, 3H), 1.77-1.48 (m, 5H).

¹³C NMR (CD₃OD): δ=168.4, 159.1, 130.0, 129.1, 125.6, 125.1, 123.0,116.9, 57.2, 48.8, 46.3, 44.5, 31.5, 25.6, 19.3, 15.4.

Example 40 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine

The title compound was prepared from the product of Example 82 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ=7.73 (d, 2H), 7.41 (d, 2H), 7.27 (d, 2H), 7.04 (d,2H), 4.68-4.65 (m, 1H), 4.10-4.07 (m, 1H), 3.90 (t, 2H), 3.77 (t, 2H),3.38-3.11 (m, 4H), 2.66 (m, 4H), 2.43 (s, 3H), 1.80-1.48 (m, 5H).

¹³C NMR (CD₃OD): δ=168.4, 168.2, 149.4, 145.7, 139.8, 129.7, 129.0,125.6, 125.1, 123.1, 116.9, 57.2, 48.8, 44.6, 42.1, 36.0, 31.4, 25.7,22.1, 21.8, 19.3, 15.4.

Example 41 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(isonipecotoyloxy)phenylalanine

The title compound was prepared from the product of Example 80 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ=8.08 (m, 1H), 7.73 (d, 2H), 7.41 (d, 2H), 7.27 (d,2H), 7.03 (d, 2H), 4.71 (m, 1H), 4.11-4.08 (m, 1H), 3.61 (t, 2H),3.47-3.38 (m, 3H), 3.31-3.11 (m, 4H), 2.43 (s, 3H), 1.77-1.47 (m, 10H).

¹³C NMR (CD₃OD): δ=168.3, 168.1, 158.8, 149.6, 145.9, 139.8, 129.5,129.0, 125.6, 125.1, 123.1, 116.9, 57.2, 48.6, 44.6, 40.6, 40.1, 36.0,31.4, 25.7, 20.9, 20.6, 19.3.

Example 42 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(pyrrolidin-1-ylcarbonyloxy)phenylalanine

The title compound was prepared from the product of Example 83 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ=8.08 (m, 1H), 7.73 (d, 2H), 7.41 (d, 2H), 7.04 (d,2H), 7.27 (d, 2H), 4.72-4.68 (m, 1H), 4.11-4.08 (m, 1H), 3.57-3.53 (t,2H), 3.43-3.28 (m, 3H), 3.25-3.06 (m, 4H), 2.43 (s, 3H), 1.99-1.80 (m,4H), 1.78-1.49 (m, 5H).

¹³C NMR (CD₃OD): δ=168.2, 158.3, 149.2, 145.8, 139.8, 129.4, 129.1,125.6, 125.1, 123.1, 116.9, 57.2, 48.7, 44.5, 41.5, 31.4, 25.7, 20.6,19.8, 19.3, 15.4.

Example 43 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(morpholin-4-ylcarbonyloxy)phenylalanine

The title compound was prepared from the product of Example 108 usingthe procedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ=7.73 (d, 2H), 7.41 (d, 2H), 7.27 (d, 2H), 7.04 (d,2H), 4.95-4.93 (m, 1H), 4.10-4.07 (m, 1H), 3.71-3.65 (m, 6H), 3.50 (t,2H), 3.40-3.10 (m, 4H), 2.43 (s, 3H), 1.78-1.48 (m, 4H).

¹³C NMR (CD₃OD): δ=168.4, 168.2, 149.6, 145.7, 139.8, 129.1, 125.6,125.1, 123.1, 116.8, 61.5, 57.2, 44.5, 36.0, 31.4, 25.6, 19.3, 15.4.

Example 44 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanineNeopentyl Ester

Titanium isopropoxide (0.3 equivalents) was added to Tos-Pro-Tyr ethylester (1 equivalent) and an excess of neopentyl alcohol. The mixture washeated to reflux under an argon atmosphere overnight. Excess neopentylalcohol was removed under reduced pressure and the residue purified byflash column chromatography (silica, hexane: EtOAc 2:1) to give theneopentyl ester a white solid (0.9 g, 85%). The title compound wasprepared following the procedure described in Example 4.

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHz) δ=8.29 (d, 1H, J=7.91 Hz); 7.68 (d, 2H, J=8.45Hz); 7.40 (d, 2H, J=8.34 Hz); 7.24 (d, 2H, J=8.57Hz); 7.00 (d, 2H,J=8.57 Hz); 4.56 (m, 1H); 4.07 (m, 1H); 3.73 (s, 2H); 3.55 (br s, 2H);3.40 (m, 3H); 3.10 (m, 3H); 2.40 (s, 3H); 2.35 (br s, 4H); 2.20 (s, 3H);1.55 (m, 3H); 1.37 (m, 1H); 0.85 (s, 9H).

Example 45 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineNeopentyl Ester

Titanium isopropoxide (0.3 equivalents) was added to Tos-Pro-Tyr ethylester (1 equivalent) and an excess of neopentyl alcohol. The mixture washeated to reflux under an argon atmosphere overnight. Excess neopentylalcohol was removed under reduced pressure and the residue purified byflash column chromatography (silica, hexane: EtOAc 2:1) to give theneopentyl ester a white solid (0.9 g, 85%). The title compound wasprepared following the procedure described in Example 2.

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHz) δ=8.28 (d, 1H, J=8.13 Hz); 7.68 (d, 2H, J=8.4Hz); 7.40 (d, 2H, J=7.9 Hz); 7.23 (d, 2H, J=8.56Hz), 6.99 (d, 2H, J=8.35Hz); 4.57 (m, 3H); 2.40 (s, 3H); 1.55 (m, 3H); 1.38 (m, 1H); 0.85 (s,9H).

Example 46 Synthesis of2-(Saccharin-2-yl)propionoyl-L4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure described inPreparative Example A and Example 4.

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHz) (1:1 mixture of diastereomers) δ=8.31 (m, 1H);8.26 (m, 1H); 8.03 (m, 3H); 7.20 (m, 2H); 7.00 (m, 2H); 4.73 (m, 1H);4.30 (m, 1H); 3.58 (br s, 2H); 3.40 (br s, 2H); 3.02 (m, 1H); 2.95 (m,1H); 2.35 (br s, 4H); 2.20 (s, 3H); 2.75 and 2.65 (two d, 3H); 1.35 and1.32 (two s, 9H).

Example 47 Synthesis of2-(Saccharin-2-yl)propionoyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Preparative ExampleA using the procedure described in Method 11.

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHz) (1:1 mixture of diastereomers) δ=12.75 (br s,1H); 8.28 (m, 2H); 8.05 (m, 3H); 7.20 (m, 2H); 7.00 and 9.95 (two d,2H); 4.75 (m, 1H); 4.40 (m, 1H); 3.10 (m, 1H); 3.05 (s, 3H); 2.95 (m,1H); 2.90 (s, 3H); 2.75 and 2.60 (two d, 3H).

Example 48 Synthesis ofN-(Toluene-4-sulfonyl)-L-N-methylalanyl-L4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure for thesynthesis of Example 2 with the substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.68 (d, 2H), 7.31 (d, 2H), 7.17 (d, 2H), 7.04 (d,2H), 6.87 (d, 2H), 4.67 (m, 1H), 4.48 (q, 1H), 3.09 (s, 3H), 3.00 (s,3H), 3.14-2.92 (m, 2H), 2.46 (s, 3H), 2.43 (s, 3H), 1.45 (s, 9H), 0.92(d, 3H).

¹³C NMR (CDCl₃): δ=170.2, 169.9, 154.9, 150.6, 143.9, 135.6, 133.2,130.2, 130.0, 127.3, 121.9, 82.5, 55.5, 53.7, 37.2, 36.6, 36.4, 29.7,27.8, 21.4, 11.5.

Example 49 Synthesis ofN-(Toluene-4-sulfonyl)-L-(thiamorpholin-3-carbonyl)-L4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

L-Thiamorpholine-3-carboxylic acid was prepared by the method of Larssonand Carlson (Acta Chemica Scan. 1994, 48, 517-525).N-(Toluene-4-sulfonyl)-L-thiamorpholine-3-carboxylic acid was preparedusing the procedure described in Method 1. The title compound wasprepared following the procedure for the synthesis of Example 2 withsubstitution of appropriate starting materials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.69 (d, 2H), 7.31 (d, 2H), 7.16 (d, 2H), 6.98 (d,2H), 6.86 (d, 1H), 4.71 (m, 1H), 4.62 (m, 1H), 3.94 (m, 1H), 3.31 (m,1H), 3.09 (m, 4H), 2.98 (s, 3H), 2.67 (m, 1H), 2.50 (m, 1H), 2.40 (s,3H), 2.31 (m, 1H), 2.10 (m, 1H), 1.49 (s, 9H).

³C NMR (CDCl₃): δ=169.9, 167.4, 154.8, 150.6, 144.2, 136.8, 132.8,130.4, 130.2, 127.3, 121.8, 82.6, 55.2, 54.0, 43.3, 36.5, 36.3, 27.8,25.2, 24.6, 21.4.

Example 50 Synthesis ofN-(Toluene-4-sulfonyl)sarcosyl-L-4-(1,1-dioxothiomorphotin-4-ylcarbonyloxy)phenylalanine

The title compound was prepared from the product of Example 121 usingthe procedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ=7.67 (d, 2H), 7.40 (d, 2H), 7.27 (d, 2H), 7.09 (d,2H), 4.61 (m, 1H), 4.12 (m, 2H), 3.99 (m, 2H), 3.60 (m, 2H), 3.23 (m,8H), 2.58 (s, 3H), 2.42 (s, 3H).

¹³C NMR (CD₃OD): δ=174.2, 170.3, 155.0, 151.6, 145.6, 136.1, 135.2,131.5, 131.1, 128.9, 123.0, 54.6, 54.0, 52.4, 52.2, 44.4, 44.0, 37.4,36.8, 21.4.

Example 51 Synthesis ofN-(Toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared from the product of Example 49 followingthe procedure described by Larsson and Carlson (Acta Chemica Scan. 1994,48, 522).

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.76 (d, 2H), 7.37 (d, 2H), 7.08 (d, 2H), 6.98 (d,2H), 6.56 (d, 1H), 4.95 (m, 1H), 4.62 (m, 1H), 3.99 (m, 2H), 3.25 (m,1H), 3.07 (s, 3H), 2.97 (m, 8H), 2.44 (s, 3H), 1.48 (s, 9H).

¹³C NMR (CDCl₃): δ=170.0, 164.8, 154.9, 150.7, 145.4, 135.3, 132.6,130.7, 130.3, 127.5, 122.3, 82.8, 56.1, 53.6, 49.5, 48.6, 41.6, 36.6,36.4, 27.9, 21.6.

Example 52 Synthesis ofN-(Toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(morpholin-4-ylcarbonyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 71.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.75 (d, 2H), 7.36 (d, 2H), 7.12 (d, 2H), 6.98 (d,2H), 6.58 (d, 1,1H), 4.93 (m, 1H),-4.63 (m, 1H), 4.09 (m, 2H), 3.72 (m,4H), 3.63 (m, 2H), 3.51 (m, 2H), 3.24 (m, 1H), 2.96 (m, 4H), 2.43 (s,3H), 1.46 (s, 9H).

¹³C NMR (CDCl₃): δ=170.0, 164.8, 153.7, 150.4, 145.4, 135.2, 132.9,130.7, 130.4, 127.5, 122.1, 82.9, 66.4, 56.1, 53.6, 49.4, 48.5, 44.7,43.9, 41.6, 36.3, 27.8, 21.6.

Example 53 Synthesis ofN-(Toluene-4-sulfonyl)-L-N-methylalanyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 48 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.68 (d, 2H), 7.31 (d, 2H), 7.20 (d, 2H), 7.11-7.04(m, 3H), 6.35 (br s, 1H), 4.81 (m, 1H), 4.52 (q, 1H), 3.35-2.98 (m, 2H),3.09 (s, 3H), 3.00 (s, 3H), 2.45 (s, 3H), 2.43 (s, 3H), 0.91 (d, 3H).

¹³C NMR (CDCl₃): δ=173.7, 170.8, 155.2, 150.6, 144.0, 135.4, 133.2,130.2, 130.0, 127.3, 122.1, 55.5, 53.2, 36.6, 36.5, 36.4, 29.8, 21.4,11.6.

Example 54 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

L-Thiamorpholine-3-carboxylic acid was prepared by the method of Larssonand Carlson (Acta Chemica Scan. 1994, 48, 517-525).N-(Toluene-4-sulfonyl)-L-thiamorpholine-3-carboxylic acid was preparedusing the procedure described in Method 1.

The title compound was then prepared following the procedure for thesynthesis of Example 2.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.87-7.82 (m, 2H), 7.20 (t, 2H), 7.16 (d, 2H), 7.00(d, 2H), 6.76 (d, 1H), 4.74 (t, 1H), 4.65 (q, 1H), 3.92 (d, 1H), 3.32(dd, 1H), 3.17-3.00 (m, 2H), 3.09 (s, 3H), 2.99 (s, 3H), 2.76-2.66 (m,1H), 2.62 (dd, 1H), 2.46 (dt, 1H), 2.22 (d, 1H), 1.49 (s, 9H).

¹³C NMR (CDCl₃): δ=70.0, 167.2, 165.5, 154.8-150.7-135.8-132.7, 130.5,130.1, 121.9, 116.9, 82.8, 55.3, 53.9, 43.4, 36.6, 36.4, 36.3, 27.9,25.8, 25.0.

Example 55 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared from the product of Example 54 followingthe procedure described by Larsson and Carlson (Acta Chemica Scan. 1994,48, 522).

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.92-7.88 (m, 2H), 7.24 (t, 2H), 7.09 (d, 2H), 6.97(d, 2H), 6.41 (d, 1H), 4.96 (d, 1H), 4.62 (d, 1H), 4.03 (d, 1H), 3.26(dd, 1H), 3.13-2.92 (m, 6H), 3.09 (s, 3H), 2.97 (s, 3H), 1.49 (s, 9H).

¹³C NMR (CDCl₃): δ=170.1, 165.9, 164.5, 154.9, 150.7, 134.0, 132.4,130.5, 130.4, 122.2, 117.3, 83.0, 56.1, 53.4, 50.0, 49.1, 41.7, 36.6,36.3, 36.1, 27.9.

Example 56 Synthesis ofN-(Pyridine-3-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

N-Benzyl-L-proline was coupled to L-tyrosine t-butyl ester using theprocedure described in Method 12.N-Benzyl-L-prolyl-L-(N,N-dimethylcarbamyloxy)phenyl-alanine t-butylester was prepared following the procedure described for the preparationof Example 2 . L-Prolyl-L-(4-N,N-dimethylcarbamyloxy)phenylalaninet-butyl ester was prepared from the product of the previous reactionusing the procedure described in Method 4. The title compound wasprepared using the procedure described for the preparation of 3-pyridinesulfonyl chloride (see Crowell et al., J. Med. Chem., 1989, 32,2436-2442) and the product of the last reaction.

NMR data was as follows:

¹H NMR (CDCl₃): δ=9.95 (d, 1H), 8.83 (dd, 1H), 8.14-8.10 (m, 1H),7.51-7.47 (m, 1H), 7.16-7.13 (m, 3H), 7.02-6.99 (m, 2H), 4.72-4.69 (m,1H), 4.09-4.06 (m, 1H), 3.41-3.39 (m, 1H), 3.23-3.17 (m, 1H), 3.13-2.98(m, 1H), 3.07 (s, 3H), 2.97 (s, 3H), 2.04 (m, 1H), 1.59-1.47 (m, 3H),1.45 (s, 9H)

¹³C NMR (CDCl₃): δ=170.1, 169.9, 154.8, 153.9, 150.5, 148.4, 135.5,133.0, 130.1, 123.9, 121.6, 82.6, 52.2, 53.6, 49.5, 37.1, 36.5, 36.3,29.9, 27.8, 24.0.

Preparative Example B Synthesis ofN-(Pyrimidine-2-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared by substituting 2-pyrimidine sulfonylchloride (see Skulnick et al., J. Med. Chem., 1997, 40, 1149-1164) andfollowing the method for the preparation of Example 56.

NMR data was as follows:

¹H NMR (CDCl₃): δ=8.28 (d, 2H), 7.39 (d, 1H), 7.02 (d, 2H), 6.88 (d,2H), 6.54 (m, 1H), 4.76-4.69 (m, 1H), 4.57-4.55 (m, 1H), 3.64 (m, 1H),3.55-3.52 (m, 1H), 3.09-3.03 (m, 1H), 3.08 (s, 3H), 2.99-2.95 (m, 1H),2.98 (s, 3H), 2.32 (m, 1H), 2.01-1.97 (m, 3H), 1.37 (s, 9H).

¹³C NMR (CDCl₃): δ=172.1, 170.4, 160.6, 157.7, 154.8, 150.3, 133.0,130.1, 121.3, 110.5, 82.0, 60.7, 53.3, 47.5, 37.1, 36.5, 36.3, 28.9,27.7, 24.1.

Example 57 Synthesis ofN-(4-Nitrobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 35 using theprocedure described in method 11.

NMR data was as follows:

¹H NMR (CDCl₃): δ=8.36 (d, 2H), 8.02 (d, 2H), 7.42 (d, 1H), 7.20 (d,2H), 7.01 (d, 2H), 4.86 (m, 1H), 4.18-4.15 (m, 1H), 3.46-3.43 (m, 1H),3.32-3.26 (m, 1H), 3.19-3.11 (m, 2H), 3.09 (s, 3H), 3.01 (s, 3H), 1.91(m, 1H), 1.65-1.54 (m, 3H).

¹³C NMR (CDCl₃): δ=172.9, 171.7, 155.5, 150.4, 150.4, 142.1, 133.2,130.5, 129.1, 124.6, 121.8, 61.9, 52.9, 49.6, 36.6, 36.3, 36.3, 30.6,24.1.

Example 58 Synthesis ofN-(4-Cyanobenzenesulfonyl)-L-prolyl-L-4-(NN-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.94 (d, 2H), 7.82 (d, 2H), 7.13 (d, 2H), 7.05-6.99(m, 3H), 4.71-4.66 (m, 1H), 4.12-4.09 (m, 1H), 3.36-3.35 (m, 1H),3.22-3.11 (m, 2H), 3.07 (s, 3H), 3.06-3.01 (m, 1H), 2.97 (s, 3H), 2.05(m, 1H), 1.63-1.37 (m, 3H), 1.46 (s, 9H).

¹³C NMR (CDCl₃): δ=170.1, 169.9, 154.8, 150.6, 140.8, 133.1, 132.9,130.2, 128.4, 121.7, 117.1, 116.9, 82.7, 62.2, 53.4, 49.4, 37.0, 36.5,36.3, 30.1, 27.8, 24.1.

Example 59 Synthesis ofN-(Toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L4-(N,N-dimethylaminosulfonyloxy)phenylalanine

The title compound was prepared from the product of Example 36 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ=7.79 (m, 2H), 7.44 (m, 2H), 7.27 (m, 2H), 7.17 (m,2H), 5.21 (m, 1H), 4.64 (m, 1H), 4.14 (m, 1H), 3.61 (m, 2H)? 3.24 (m,2H), 3.08 (m, 2H), 2.89 (s, 6H), 2.80 (m, 2H), 2.43 (s, 3H).

¹³C NMR (CD₃OD): δ=173.9, 168.1, 168.0, 150.8, 150.8, 146.7, 146.5,137.6, 137.5, 137.1, 136.9, 132.2, 132.1, 131.7, 131.6, 128.8, 123.3,123.1, 57.3, 54.8, 51.0, 50.8, 50.5, 47.9, 47.8, 43.2, 43.0, 39.0, 39.0,37.4, 37.0, 21.5.

Example 60 Synthesis ofN-(Toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 51 using theprocedure described in Method 1.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.79 (d, 2H), 7.43 (d, 2H), 7.20 (d, 2H), 7.00 (d,2H), 5.21 (m, 1H), 4.65 (m, 1H), 4.12 (m, 1H), 3.75 (m, 1H), 3.29 (m,3H), 3.08 (s, 3H), 3.00 (m, 1H), 3.00 (m, 1H), 2.97 (s, 3H), 2.80 (m,3H), 2.44 (s, 3H).

¹³C NMR (CDCl₃): δ=165.1, 159.0, 147.9, 143.1, 137.6, 128.6, 126.1,122.7, 122.6, 119.8, 114.3, 48.3, 45.8, 41.6, 34.0, 28.0, 27.8, 27.7,12.5.

Example 61 Synthesis ofN-(Toluene-4-sulfonyl)-L-(1,1-dioxo)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared fromN-(toluene-4-sulfonyl)-L-thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester, prepared as per the examples herein, following theprocedure described by by Larsson and Carlson (Acta Chemica Scan. 1994,48, 522).

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.77 (d, 2H), 7.38 (d, 2H), 7.18 (m, 3H), 7.09 (d,2H), 4.83-4.57 (m, 3.H), 3.77-3.60 (m, 2H), 3.36-3.23 (m, 1H), 3.15-3.00(m, 7H), 2.85-2.73 (m, 1H), 2.46 (s, 3H), 1.50 (s, 9H).

Example 62 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-prolyl-L-4-(NN-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure outlined for thepreparation of Example 2 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.96 (d, 2H), 7.80 (d, 2H), 7.26-7.13 (m, 3H), 7.01(d, 2H), 4.72-4.70 (m, 1H), 4.11-4.08 (m, 1H), 3.40-3.37 (m, 1H),3.25-3.10 (m, 2H), 3.07 (s, 3H), 3.04-3.02 (m, 1H), 2.98 (s, 3H), 2.06(m, 1H), 2.06-2.04 (m, 1H), 1.61-1.52 (m, 3H), 1.46 (s, 9H).

¹³C NMR (CDCl₃): δ=170.3, 169.9, 154.9, 150.6, 139.9, 134.9, 133.1,130.2, 128.4, 126.5, 121.7, 82.7, 62.3, 5.35, 49.6, 37.2, 36.6, 36.3,30.0, 27.8 24.1.

Example 63 Synthesis ofN-(1-Methylpyrazolyl4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 117 usingthe procedure described in Method 11.

NMR data was as follows:

¹H NMR (CDCl₃): δ=8.84 (br s, 1H), 7.93 (s, 1H), 7.79 (s, 1H), 7.68-7.65(m, 1H), 7.18 (d, 2H), 6.99 (d, 2H), 4.88-4.81 (m, 1H), 4.08-4.06 (m,1H), 3.92 (s, 3H), 3.45-3.40 (m, 1H), 3.34-3.27 (m, 1H), 3.11-3.01 (m,5H), 2.97 (s, 3H), 1.82 (m, 1H), 1.66-1.57 (m, 2H), 1.45 (m, 1H).

¹³C NMR (CDCl₃): δ=173.1, 172.9, 159.1, 158.6, 150.4, 138.8, 133.4,133.2, 130.3, 121.9, 117.3, 62.0, 53.1, 49.7, 39.4, 36.6, 36.5, 36.4,30.4, 23.9.

Example 64 Synthesis ofN-(Toluene4-sulfonyl)-L-(1,1-dioxo)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 61 using theprocedure described in Method 11.

NMR data was as follows:

1H NMR (CDCl₃): δ=8.34 (d, 1H), 7.70 (d, 2H), 7.33 (d, 2H), 7.14 (d,2H), 7.01 (d, 2H), 5.07 (m, 1H), 4.93 (m, 1H), 4.43 (d, 1H), 4.01 (d,1H), 3.68 (m, 1H), 3.37 (m, 1H), 3.17 (s, 3H), 3.14 (m, 1H), 3.09 (s,3H), 2.54 (m, 1H), 2.43 (s, 3H).

¹³C NMR (CDCl₃): δ=171.5, 166.4, 156.4, 150.5, 145.5, 134.2, 134.1,131.4, 130.3, 128.1, 121.8, 64.3, 59.2, 53.7, 50.5, 36.9, 36.5, 35.8,21.6.

Example 65 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 84 using theprocedure described in Method 1.

NMR data was as follows:

1H NMR (CDCl₃): δ=7.83 (m, 2H), 7.73 (d, 1H), 7.16 (m, 4H), 6.99 (d,2H), 5.57 (br s, 1H), 4.87 (m, 1H), 4.76 (m, 1H), 4.53 (d, 1H), 4.10 (d,1H), 3.34 (m, 1H), 3.22 (d, 2H), 3.12 (s, 3H), 3.04 (s, 3H), 2.43 (m,1H).

¹³C NMR (CDCl₃): δ=172.1, 168.7, 155.7, 150.5, 133.6, 133.1, 130.8,130.7, 121.7, 116.9, 116.6, 65.3, 53.3, 51.3, 36.8, 36.4, 36.1, 33.4.

Example 66 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-(5,5-dimethyl)-thiaprolyl-L4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 with the substitution of appropriatestarting materials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.91 (m, 2H), 7.26 (m, 4H), 7.02 (d, 2H), 6.96 (d,1H), 4.75 (m, 1H), 4.55 (d, 1H), 4.42 (d, 1H), 3.86 (s, 1H), 3.08 (s,3H), 3.05 (m, 2H), 3.00 (s, 3H), 1.43 (s, 9H), 1.17 (s, 3H), 1.16 (s,3H).

¹³C NMR (CDCl₃): δ=169.9, 168.1, 167.6, 164.2, 154.9, 150.6, 133.1,132.2, 131.0, 130.9, 130.4, 121.7, 116.9, 116.6, 82.7, 73.5, 54.7, 53.7,50.5, 37.8, 36.6, 36.4, 29.1, 27.8, 23.8.

Example 67 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(1,1-dioxothiomorpholin-4-ylcarbonyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 68.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.91-7.87 (m, 2H), 7.27-7.25 (m, 2H), 7.15 (d, 2H),6.51 (d, 1H), 4.93-4.90 (m, 1H), 4.64 -4.58 (m, 1H), 4.14-3.99 (m, 7H),3.28-2.90 (m, 10H), 1.47 (s, 9H).

¹³C NMR (CDCl₃): δ=170.1, 167.6, 164.5, .153.1, 149.8, 133.9, 133.4,130.7, 130.5, 121.7, 117.4, 117.1, 83.1, 56.1, 53.4, 51.6, 49.9, 48.9,43.1, 41.6, 36.2, 27.8.

Example 68 Synthesis ofN-(Toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L4-(1,1-dioxothiomorpholin-4-ylcarbonyloxy)phenylalaninetert-Butyl Ester

Substituting thiomorpholine for N-methylpiperazine, and following themethod for the preparation of Example 4 and oxidation of the sulfurgroup in the thiomorpholino ring per by Larsson and Carlson (ActaChemica Scan. 1994, 48, 522) gave the title compound as a white solid.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.75 (d, 2H), 7.35 (d, 2H), 7.17 (d, 2H), 6.99 (d,2H), 6.65 (d, 1H), 4.92-4.90 (m, 1H), 4.63-4.60 (m, 1 H), 4.15-3.95 (m,7H), 3.30-3.23 (m, 1 H), 3.14 (t, 4H), 3.07-2.80 (m, 6H), 2.45 (s, 3H),1.48 (s, 9H).

¹³C NMR (CDCl₃): δ=169.9, 164.8, 153.1, 149.8, 145.5, 135.1, 133.6,130.7, 127.5, 121.8, 82.9, 60.3, 56.1, 53.7, 51.8, 49.3, 48.4, 43.1,42.7, 41.5, 36.3, 27.8, 21.5.

Example 69 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure described inExample 37 and substitution of appropriate starting materials.

NMR data was as follows:

¹³H NMR (CDCl₃): δ=7.88-7.83 (m, 2H), 7.26-7.15 (m, 5H), 7.01 (d, 2H),4.74-4.67 (m, 1H), 4.08-4.05 (m, 1H), 3.91-3.80 (m, 4H), 3.41-3.35 (m,1H), 3.24-3.00 (m,3H),.2.70-2.65 (t, 4H), 2.06-2.04 (m, 1H). 1.60-1.46(m, 12H).

¹³C NMR (CDCl₃): δ=170.5, 169.8, 153.4, 150.2, 133.5, 130.7, 130.5,130.3, 121.6, 116.8, 116.5, 82.6, 62.2, 53.6, 49.6, 47.0, 46.4, 37.2,29.8, 27.8, 27.3, 27.0, 24.1.

Example 70 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-(5,5-dimethyl)-thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 66 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.90 (m, 2H), 7.30-7.14 (m, 5H), 7.02 (d, 2H), 5.83(br s, 1H), 4.90 (m, 1H), 4.57 (d, 1H), 4.40 (d, 1H), 3.96 (s, 1H), 3.09(s, 3H), 3.28-3.02 (m, 2H), 3.00 (s, 3H), 1.13 (s, 6H).

¹³C NMR (CDCl₃): δ=173.2, 169.2, 164.2, 163.9, 155.3, 150.6, 133.1,132.0, 131.0, 130.9, 130.6, 122.0, 417.0, 116.7, 73.3, 54.6, 53.3, 50.5,37.0, 36.7, 36.4, 29.0, 23.7.

Example 71 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-(1,1-dioxothiamorphotin-3-carbonyl)-L-4-(morpholin-4-ylcarbonyloxy)phenylalaninetert-Butyl Ester

Substituting 4-morpholinecarbamyl chloride for dimethylcarbamylchloride, and following the methods for the preparation of Example 2,gave the title compound as a white solid.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.91-7.87 (m, 2H), 7.26-7.20 (m, 2H), 7.11 (d, 2H),6.98 (d, 2H), 6.43 (d, 1H), 4.95-4.92 (m, 1H), 4.62-4.60 (m, 1H),4.05-4.00 (m, 2H), 3.74 (t, 4H), 3.66-3.52 (m, 4H), 3.30-2.92 (m, 6H),1.48 (s, 9H).

¹³C NMR (CDCl₃): δ=170.1, 164.5, 150.4, 134.6, 132.7, 130.5, 122.0,117.4, 117.1, 83.1, 66.5, 56.1, 53.4, 49.9, 49.0, 44.7, 44.0, 41.6,36.2, 27.8.

Example 72 Synthesis ofN-(4-Trifluoromethoxybenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.89 (d, 2H), 7.35 (d, 2H), 7.25-7.13 (m, 3H), 7.01(d, 2H), 4.70 (m, 1H), 4.09-4.06 (m, 1H), 3.39-3.36 (m, 1H), 3.24-3.01(m, 5H), 2.98 (s, 3H), 2.05 (m, 1H), 1.62-1.47 (m, 3H), 1.46 (s, 9H).

¹³C NMR (CDCl₃): δ=170.4, 169.9, 154.9, 152.7, 150.6, 134.6, 113.2,130.2, 130.1, 121.7, 120.2, 82.7, 62.2, 53.6, 49.6, 37.2, 36.6, 36.3,29.9, 27.8, 24.1.

Example 73 Synthesis ofN-(Toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

Following the method for the preparation of Example 2 and oxidation ofthe sulfur group in the thiomorpholino ring per by Larsson and Carlson(Acta Chemica Scan. 1994, 48, 522) gave the title compound.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.70 (d, 2H), 7.31 (d, 2H), 7.04 (d, 2H), 6.93 (d,2H), 6.59 (d, 1H), 5.01 (m, 2H), 4.65 (m, 1H), 4.01 (d, 1H), 3.90 (d,1H), 3.25 (m, 1H), 3.00 (s, 3H), 2.82 (m, 8H), 2.37 (s, 3H), 1.22 (s,3H), 1.20 (s, 3H).

¹³C NMR (CDCl₃): δ=170.3, 165.0, 154.6, 150.5, 145.1, 135.2, 132.3,130.4, 130.0, 127.2, 122.1, 69.5, 55.9, 53.1, 49.1, 48.5, 41.4, 36.3,36.1, 35.9, 21.4.

Example 74 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-(1,1-dioxo-5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared from the product of Example 66 followingthe procedure described by Larsson and Carlson (Acta Chemica Scan. 1994,48, 522). NMR data was as follows:

¹H NMR (CDCl₃): δ=7.88 (m, 2H), 7.24 (m, 4H), 7.05 (d, 2H), 6.95 (d,1H), 4.80 (m, 1H), 4.40 (m, 2H), 4.10 (s, 1H), 3.17-3.03 (m, 2H), 3.10(s, 3H), 3.01 (s, 3H), 1.47 (s, 9H), 1.36 (s, 3H), 1.11 (s, 3H).

¹³C NMR (CDCl₃): δ=169.8, 168.6, 166.0, 154.5, 150.8, 139.7, 133.0,131.5, 131.4, 130.3, 122.0, 117.1, 116.8, 83.0, 68.0, 60.9, 59.3, 53.8,37.4, 36.6, 36.4, 27.8, 18.9, 18.8.

Example 75 Synthesis ofN-(Toluene-4-sulfonyl)-L-(1,1-dioxo-5,5-dimethyl)thiaprolyi-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared from the product of Example 11 followingthe procedure described by Larsson and Carlson (Acta Chemica Scan. 1994,48, 522).

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.75 (d, 2H), 7.38 (d, 2H), 7.21 (d, 2H), 7.03 (m,3H), 5.08 (m, 1H), 4.89 (m, 1H), 4.38 (m, 2H), 4.10 (s, 1H), 3.22-3.04(m, 2H), 3.10 (s, 3H), 3.00 (s, 3H), 2.43 (s, 3H), 1.26 (m, 9H), 1.09(s, 3H).

¹C NMR (CDCl₃): δ=170.3, 166.3, 150.8, 145.9, 132.8, 131.9, 130.3,128.6, 122.0, 69.8, 68.0, 60.9, 59.4, 53.4, 37.4, 36.6, 36.4, 21.6,21.5, 19.2, 18.6.

Example 76 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 74 using theprocedure described in Method 11.

NMR data was as follows:

¹³C NMR (CDCl₃): δ=171.7, 167.9, 137.3, 164.5, 155.9,150.4, 133.6,131.8, 131.3, 131.2, 130.8,121.9,117.1, 116.8,67.8,60.9,59.9,53.8,36.8,36.6,36.0, 19.1, 19.0.

Example 77 Synthesis ofN-(Pyrimidine-2-sulfonyl)-L-prolyl-L-4-(NN-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Preparative ExampleB using the procedure described in Method 11.

NMR data was as follows:

¹H NMR (CDCl₃): δ=8.45 (br m, 2H), 8.22 (br s, 1H), 7.55 (d, 1H), 7.11(d, 2H), 6.95 (d, 2H), 6.81 (m, 1H), 4.80-4.74 (m, 2H), 3.70 (m, 1H),3.55 (m, 1H), 3.20-3.08 (m, 4H), 2.98 (s, 3H), 2.89-2.76 (m, 1H),2.13-1.96 (m, 3H), 1.60 (m, 1H).

¹³C NMR (CDCl₃): δ=190.0, 173.6, 171.0, 155.2, 153.9,150.6, 133.2,130.1, 121.9, 110.3, 62.0, 55.1, 48.2, 36.6, 36.6, 36.3, 30.2, 23.4.

Example 78 Synthesis ofN-(Toluene-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalaninetert-Butyl Ester

Following the method for the preparation of Example 4 and oxidation ofthe sulfur group in the thiamorpholino ring per Larsson and Carlson(Acta Chemica Scan. 1994, 48, 522) gave the title compound.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.76 (d, 2H), 7.37 (d, 2H), 7.12 (d, 2H), 6.96 (d,2H), 6.57 (d, 1H), 4.95 (m, 1H), 4.62 (m, 1H), 4.03 (m, 2H), 3.67 (m,4H), 3.25 (m, 1H), 2,89 (m, 4H), 2.45 (m, 6H), 2.35 (s, 3H), 1.48 (s,9H).

¹³C NMR (CDCl₃): δ=170.0, 164.8, 153.7, 150.5, 145.4, 135.3, 132.8,130.7, 130.4, 127.5, 122.2, 82.9, 56.2, 54.6, 54.5, 53.6, 49.5, 48.6,46.0, 44.2, 43.7, 41.6, 36.3, 27.9, 21.6.

Example 79 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-(1,1-dioxo)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 85 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CDCl₃): δ=4.98, (m, 1H), 4.90 (m,--lH-), 4.44 (d, 1H), 4.03(d,-1-H), 3.67 (m, 1H), 3.37(m, 1H), 3.25-3.02 (m, 1H), 3.20 (s, 3H),3.11 (s, 3H), 2.68 (m, 1H).

¹³C NMR (CDCl₃): δ=171.7, 167.9, 166.3, 164.4, 157.0, 156.4, 150.5,139.6, 134.0, 133.1, 131.3, 131.1, 130.9, 121.9, 117.2, 116.9, 64.1,58.8, 53.7, 50.6, 36.9, 36.5, 35.6.

Example 80 Synthesis ofN-(Toluene4-sulfonyl)-L-prolyl-L-4-(isonipecotoyloxy)phenylalaninetert-Butyl Ester

Substituting piperazine for N-methylpiperazine, and following themethods for the preparation of Example 4, gave the title compound as awhite solid.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.70 (d, 2H), 7.32-7.26 (m, 2H), 7.14 (d, 2H), 7.01(d, 2H), 4.72-4.68 (m, 1H), 4.07-4.05 (m, 1H), 3.60-3.49 (m, 4H),3.37-3.31 (m, 1H), 3.22-2.98 (m, 3H), 2.42 (s, 3H), 2.02 (m, 2H),1.61-1.55 (m, 6H), 1.50-1.45 (m, 13H).

¹³C NMR (CDCl₃): δ=177.3, 170.7, 169.8, 150.6, 144.3, 133.1, 130.1,129.9, 127.9, 121.6, 110.8, 82.5, 62.2, 57.2, 53.7, 49.5, 44.9, 37.2,29.7, 27.8, 25.7, 24.1, 21.4.

Example 81 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(1,1-dioxothiomorpholin-4-ylcarbonyloxy)phenylalaninetert-Butyl Ester

The product of Example 82 was oxidized by the method of Larsson andCarlson (Acta Chemica Scan. 1994, 48, 517-525), yielding the titlecompound as a white solid.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.69 (d, 2H), 7.33-7.29 (m, 3H), 7.20 (d, 2H), 7.00(d, 2H), 4.71-4.66 (m, 1H), 4.13-4.04 (m, 5H), 3.37-3.32 (m, 1H),3.21-3.00 (m, 7H), 2.41 (s, 3H), 2.05-2.01 (m, 1H), 1.52-1.44 (m, 12H).

¹³C NMR (CDCl₃): δ=170.7, 169.7, 149.8, 144.3, 134.4, 133.3, 130,6,130.0, 127.9, 121.4, 82.7, 62.4, 54.0, 52.1, 49.7, 43.2, 37.6, 29.7,28.1, 24.4, 21.7.

Example 82 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalaninetert-Butyl Ester

Substituting thiomorpholine for N-methylpiperazine, and following themethods for the preparation of Example 4, gave the title compound as awhite solid.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.70 (d, 2H), 7.31-7.26 (m, 2H), 7.16 (d, 2H), 7.00(d, 2H), 4.72-4.66 (m, 1H), 4.07-4.04 (m, 1H), 3.89-3.79 (m, 4H),3.37-3.32 (m, 1H), 3.22-2.99 (m, 3H), 2.67 (t, 4H), 2.42 (s, 3H), 2.02(m, 2H), 1.50-1.45 (m, 12H).

¹³C NMR (CDCl₃): δ=177.2, 170.7, 169.8, 153.5, 150.2, 144.3, 133.6,132.9, 130.3, 129.9, 127.9, 121.5, 82.5, 62.4, 53.7, 49.5, 47.0, 46.4,37.2, 29.6, 27.8, 27.3, 24.1, 21.4.

Example 83 Synthesis ofN-(Toluene4-sulfonyl)-L-prolyl-L4-(pyrrolidin-1-ylcarbonyloxy)phenylalaninetert-Butyl Ester

Substituting pyrrolidinecarbonyl chloride for dimethylcarbamyl chloride,and following the methods for the preparation of Example 2, gave thetitle compound as a white solid.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.71 (d, 2H), 7.32 (d, 2H), 7.15 (d, 2H), 7.04 (d,2H), 4.73-4.67 (m, 1H), 4.07-4.04 (m, 1H), 3.53 (t, 2H), 3.45 (t, 2H),3.36-3.32 (m, 1H), 3.24-2.98 (m, 3H), 2.42 (s, 3H), 2.03-1.88 (m, 5H),1.75 (s, 1H), 1.52 (1.24 (m, 12H).

¹³C NMR (CDCl₃): δ=170.7, 169.8, 153.1, 150.4, 144.3, 133.1, 130.1,129.9, 127.9, 121.6, 110.8, 99.8, 82.5, 62.2, 53.7, 49.5, 46.3, 37.2,29.7, 27.8, 25.6, 24.8, 24.0, 21.4.

Example 84 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedures described forthe preparation of Example 2 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.87 (m, 2H), 7.28-7.13 (m, 5H), 7.02 (d, 2H),4.70-4.60 (m, 2H), 4.58 (d, 1H), 4.06 (d, 1H), 3.38-3.01 (m, 3H), 3.09(s, 3H), 3.00 (s, 3H), 2.58 (m, 1H), 1.47 (s, 9H).

¹³C NMR (CDCl₃): δ=169.7, 167.8, 154.9, 150.7, 132.7, 130.9, 130.7,130.4, 121.8, 117.1, 116.8, 82.9, 65.1, 53.9, 51.4, 36.8, 36.6, 36.4,33.1, 27.9.

Example 85 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-(1,1-dioxo)thiaprolyl-L4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared from the product of Example 84 followingthe procedure oxidation procedure of Larsson and Carlson (Acta ChemicaScan. 1994, 48, 517-525).

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.90 (m, 2H), 7.30-7.04 (m, 7H), 4.83-4.58 (m, 3H),3.66 (m, 2H), 3.32-3.24 (m, 1H), 3.09-2.85 (m,2H), 3.10 (s, 3H), 3.01(s, 3H), 1.50 (s, 9H).

¹³C NMR (CDCl₃): δ=173.1, 169.8, 168.0, 165.6,154.9, 150.9, 132.6,131.1, 131.0, 130.3, 122.3, 117.3, 117.0, 83.2, 62.8, 57.8, 53.9, 49.0,36.8, 36.6, 36.4, 27.9.

Example 86 Synthesis ofN-(2,5-Dichlorothiophene-3-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.14 (d, 2H), 7.09 (s, 1H), 7.07 (d, 1H), 7.01 (d,2H), 4.73-4.66 (m, 1H), 4.32-4.28 (m, 1H), 3.42-3.17 (m, 3H), 3.08 (s,3H), 3.06-3.01 (m, 1H), 2.98 (s, 3H), 2.17-2.04 (m, 1H), 1.84-1.60 (m,2H), 1.60-1.46 (m, 1H), 1.45 (s, 9H).

¹³C NMR (CDCl₃): δ=170.2, 169.9, 154.9, 150.6, 133.4, 133.1, 131.2,130.2, 127.9, 27.0, 82.7, 62.2, 53.6, 49.3, 37.2, 36.6, 36.4, 30.1, 24.2.

Example 87 Synthesis ofN-(4-Acetamidobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=8.58 (s, 1H), 7.70-7.67 (m, 4H), 7.32 (d, 1H), 7.14(d, 2H), 7.01 (d, 2H), 4.68 (m, 1H), 3.99 (m, 1H), 3.37-3.34 (m, 1H),3.23-3.16 (m, 1H), 3.11-3.01 (m, 1H), 3.08 (s, 3H), 2.98 (s, 3H), 2.13(s, 3H), 1.97-1.94 (m, 1H), 1.55-1.47 (m, 3H), 1.44 (s, 9H).

¹³C NMR (CDCl₃): δ=171.1, 169.9, 169.4, 155.0, 150.6, 143.3, 133.3,130.2, 130.0, 128.9, 121.7, 119.4, 82.7, 62.2, 53.8, 49.6, 37.2, 36.6,36.4, 29.9, 27.8, 24.4, 24.1.

Example 88 Synthesis ofN-(4-tert-Butylbenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure outlined for thepreparation of Example 73 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.81 (d, 2H), 7.59 (d, 2H), 7.07 (d, 2H), 6.97 (d,2H), 6.46 (d, 1H), 4.95 (m, 1H), 4.62 (m, 1H), 4.06 (m, 2H), 3.23 (m,1H), 3.07 (m, 4H), 2.97 (m, 4H), 2.81 (m, 4H), 1.55 (s, 9H), 1.37 (s,9H).

¹³C NMR (CDCl₃): δ=170.0, 164.9, 158.2, 154.8, 150.6, 135.0, 132.6,130.2, 127.4, 126.9, 122.2, 82.7, 56.1, 53.5, 49.7, 48.8, 41.5, 36.5,36.3, 36.1, 35.2, 30.8, 27.8.

Example 89 Synthesis ofN-(Pyridine-3-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 56 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ=8.95 (s, 1H), 8.83 (d, 1H), 8.28-8.24 (m, 1H),7.73-7.69 (m, 1H), 7.30 (d, 2H), 7.05 (d, 2H), 4.68-4.63 (m, 1H),4.29-4.25 (m, 1H), 3.47-3.41 (m, 1H), 3.38-3.22 (m, 2H), 3.09 (s, 3H),3.06-3.02 (m, 1H), 2.96 (s, 3H), 1.92-1.66 (m, 4H).

¹³C NMR (CD₃OD): δ=174.2, 173.9, 160.6, 160.0, 156.9, 152.9, 152.0,147.9, 139.1, 136.9, 135.7, 131.6, 126.5, 123.1, 63.1, 54.8, 50.4, 37.5,36.8, 36.7, 32.2, 25.5.

Example 90 Synthesis ofN-(2-Fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

L-Thiamorpholine-3-carboxylic acid was prepared by the method of Larssonand Carlson (Acta Chemica Scan. 1994, 48, 517-525).N-(2-fluorobenzene-4-sulfonyl)-L-thiomorpholine-3-carboxylic acid wasprepared using the procedure described in Method 1.

The title compound was prepared according to the procedures set forthabove using suitable starting materials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.92 (m, 1H), 7.69 (m, 1H), 7.34 (m, 2H), 7.16 (m,2H), 6.99 (m, 2H), 6.60 (d, 1H), 5.01 (m, 1H), 4.64 (m, 1H), 4.03 (m,2H), 3.29 (m, 1H), 3.06 (m, 6H), 2.90 (m, 7H), 1.49 (d, 9H).

¹³C NMR (CDCl₃): δ=169.9, 164.8, 160.3, 156.9, 154.9, 150.7, 136.6,136.4, 132.7, 131.0, 130.3, 128.8, 126.4, 126.2, 125.1, 122.2, 118.1,117.8,82.7, 56.3,56.7,50.2,49.5, 41.8, 36.5, 36.3, 27.8.

Example 91 Synthesis of N-(3-Fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

L-Thiamorpholine-5-carboxylic acid was prepared by the method of Larssonand Carlson (Acta Chemica Scan. 1994, 48, 517-525).N-(3-fluorobenzene-4-sulfonyl)-L-thiamorpholine-5-carboxylic acid wasprepared using the procedure described in Method 1.

The title compound was prepared according to the procedures set forthabove using suitable starting materials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.66 (m, 1H), 7.58 (m, 2H), 7.34 (m, 1H), 7.07 (d,1H), 6.92 (d, 1H), 6.42 (d, 1H), 5.00 (m, 1H), 4.58 (m, 1H), 4.02 (m,2H), 3.22 (m, 1H), 3.05 (s, 3H), 2.98 (m, 6H), 1.45 (s, 9H).

¹³C NMR (CDCl₃): δ=170.0, 164.5, 164.4, 161.0, 154.9, 150.6, 140.3,140.2, 132.5, 131.9, 131.8, 130.2, 123.2, 123.1, 122.2, 121.4, 121.2,115.0, 114.7, 82.9, 56.1, 53.4, 49.9, 49.1, 41.7, 36.5, 36.3, 36.0,27.8.

Example 92 Synthesis ofN-(2,4-Difluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

L-Thiamorpholine-5-carboxylic acid was prepared by the method of Larssonand Carlson (Acta Chemica Scan. 1994, 48, 517-525).N-(2,4-difluorobenzene-4-sulfonyl)-L-thiamorpholine-5-carboxylic acidwas prepared using the procedure described in Method 1.

The title compound was prepared according to the procedures set forthabove using suitable starting materials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.93 (m, 1H), 7.15 (m, 2H), 7.04 (m, 4H), 6.53 (d,1H), 4.97 (m, 1H), 4.64 (m, 1H), 4.05 (m, 2H), 3.21 (m, 3H), 3.17 (s,3H), 2.97 (m, 5H), 1.43 (s, 9H).

¹³C NMR (CDCl₃): δ=170.0, 164.6, 154.9, 150.7, 132.6, 132.6, 130.3,122.6, 122.1, 112.6, 112.3, 107.0, 106.7, 106.3, 82.8, 56.3, 53.5, 50.5,49.8, 42.0, 36.5, 36.3, 27.8.

Example 93 Synthesis ofN-(4-Acetamidobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 87 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃0D): δ=8.05 (d, 1H), 7.78 (m, 4H), 7.26 (d, 2H), 7.02 (d,2H), 4.94 (m, 1H), 4.72-4.67 (m, 1H), 4.13-4.09 (m, 1H), 3.40-3.36 (m,1H), 3.30-3.05 (m, 3H), 3.08 (s, 3H), 2.97 (s, 3H), 2.15 (s, 3H),1.81-1.51 (m, 4H).

¹³C NMR (CD₃OD): δ=174.3, 174.2, 172.3, 156.9, 152.0, 144.9, 135.5,132.4, 131.6, 130.2, 122.9, 120.7, 63.2, 54.7, 50.6, 37.5, 36.8, 36.7,31.7, 25.4, 24.0.

Example 94 Synthesis ofN-(4-Trifluoromethoxybenzenesulfonyl)-L-prolyl-L4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 72 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ=7.93 (d, 2H), 7.48 (d, 2H), 7.28 (d, 2H), 7.03 (d,2H), 4.72- 4.68 (m, 1H), 4.17-4.13 (m, 1H), 3.45-3.42 (m, 1H), 3.28-3.11(m, 2H), 3.14-3.07 (m, 1H), 3.09 (s, 3H), 2.97 (s, 3H), 1.85-1.69 (m,3H), 1.59 (m, 1H).

¹³C NMR (CD₃OD): δ=174.2, 174.1, 157.0, 153.9, 152.0, 137.3, 135.6,131.7, 131.5, 123.0, 122.5, 121.8, 63.1, 54.7, 50.6, 37.4, 36.8, 36.6,31.9, 25.4.

Example 95 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L4-(N,N-dimethylcarbamyloxy)phenylalanine tert-Butyl Ester

The title compound was prepared using the procedure described in Example2 and substitution of appropriate starting materials.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.90 (m, 2H), 7.22 (m, 4H), 7.00 (m, 3H), 5.08 (m,1H), 4.84 (m, 1H), 4.56 (d, 1H), 4.42 (d, 1H), 3.88 (s, 1H), 3.15-2.99(m, 2H), 3.09 (s, 3H), 3.00 (s, 3H), 1.26-1.16 (m, 12H).

¹³C NMR (CDCl₃): δ=70.4, 168.2, 167.5, 164.1, 154.9,150.7, 132.8, 132.2,132.1, 131.0, 130.8, 130.3, 121.8, 116.9, 116.6, 73.5, 69.6, 54.6, 53.2,50.5, 37.6, 36.6, 36.3, 23.8, 21.6, 21.5.

Example 96 Synthesis ofN-(4-Cyanobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 58 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ=8.14 (d, 1H), 7.94-7.89 (m, 4H), 7.29 (d, 2H), 7.03(d, 2H), 4.70-4.66 (m, 1H), 4.21-4.17 (m, 1H), 3.47-3.40 (m, 1H),3.31-3.21 (m, 2H), 3.11-3.04 (m, 1H), 3.09 (s, 3H), 2.97 (s, 3H),1.87-1.72 (m, 3H), 1.70-1.61 (m, 1H).

¹³C NMR (CD₃OD): δ=174.2, 173.9, 157.0, 152.0, 142.9, 135.7, 134.5,131.7, 129.7, 123.0, 118.6, 111.8, 63.0, 54.7, 50.5, 37.4, 36.8, 36.7,32.0, 25.4.

Example 97 Synthesis ofN-(Toluene-4-sulfonyl)-L-(3,3-dimethyl)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine tert-Butyl Ester

The title compound was prepared using the procedure described for thepreparation of Example 98.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.76 (d, 1H), 7.75 (d, 1H), 7.35 (d, 1H), 7.34 (d,1H), 7.33 (d, 1H), 7.20 (d, 1H), 7.10 (d, 1H), 7.03 (d, 1H), 6.91 (d,0.5H), 6.08 (d, 0.5H), 4.86 (ddd, 0.5H), 4.77 (q, 0.5H), 3.61 -3.47 (m,2H), 3.27-3.02 (m, 3H), 3.09 (s, 3H), 3.00 (s, 3H), 2.45 (s, 1.5H), 2.43(s, 1.5H), 1.75-1.68 (m, 0.5H), 1.61-1.51 (m, 0.5H), 1.45 (s, 4.5H),1.40 (s, 4.5H), 1.48-1.25 (m, 3H); 0.95 (s, 1.5H), 0.80 (s, 1.5H); 0.61(s, 1.5H).

¹³C NMR (CDCl₃): δ=170.4, 170.1, 170.0, 169.6, 155.0, 154.9, 150.7,150.6, 144.3, 144.2, 133.4, 133.1, 132.8, 132.6, 130.7, 130.2, 129.9,129.8, 128.0, 121.8, 121.7, 82.6, 82.2, 71.5, 71.2, 53.6, 52.7, 47.3,47.2, 42.7, 42.5, 38.2, 38.1, 37.7, 37.5, 36.6, 36.3, 27.8, 27.8, 27.2,23.4, 23.2, 21.5.

Example 98 Synthesis ofN-(Toluene-4-sulfonyl)-L-(3,3-dimethyl)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

3,3-Dimethyl proline (see Sharma and Lubell, J. Org. Chem. 1996, 61,202-209) was N-tosylated using the procedure described in Method 1. Thetitle compound was then prepared following the procedure described forthe preparation of Example 2.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.76 (d, 1H), 7.74 (d, 1H), 7.36 (d, 1H), 7.33 (d,2H), 7.19 (d, 1H), 7.10 (d, 1H), 7.03 (d, 1H), 6.91 (d, 0.5H), 6.89 (d,0.5H), 5.06 (sept., 0.5H), 4.96 (sept., 0.5H), 4.98-4.83 (m, 1H),3.59-3.48 (m, 2H), 3.31-3.03 (m, 3H), 3.09 (s, 3H), 3.00 (s, 3H), 2.45(s, 1.5H), 2.43 (s, 1.5H), 1.75-1.66 (m, 0.5), 1.62-1.52 (m, 0.5H),1.34-1.22 (m, 3H), 1.27 (s, 1.5H), 1.25 (s, 1.5H), 1.22 (s, 1.5H), 1.20(s, 1.5H), 0.95 (s, 1.5H), 0.78 (s, 1.5H), 0.60 (s, 1.5H), 0.57 (s,1.5H).

¹³C NMR (CDCl₃): δ=170.8, 170.6, 170.0, 169.7, 154.9, 150.8, 150.6,144.4, 144.2, 133.2, 132.5, 132.5, 130.7, 130.2, 129.9, 129.8, 128.0,122.0, 121.8, 71.5, 17.2, 69.5, 69.3, 53.0, 52.2, 47.3, 47.2, 42.8,42.5, 38.2, 38.1, 37.6, 37.2, 36.6, 36.3, 27.1, 23.4, 23.2, 21.6, 21.6,21.5, 21.5.

Other compounds prepared by the methods described above include thoseset forth in Examples 99-137. In addition, Examples 101, 109, 111, 117,132 and 137 are exemplified as follows:

Example 101 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-4-(NN-dimethylcarbamyloxy)-L-phenylalanineIsopropyl Ester

The title compound was prepared following the procedure outlined for thepreparation of Example 2.

NMR data was as follows:

¹H NMR (CD₃)₂SO: δ=8.28 (d, 1H), 7.70 (d, 2H), 7.41 (d, 2H), 7.23 (d,2H), 6.99 (d, 2H), 4.86 (sept, 1H), 4.47 (m, 10H), 4.40.(m, 1H), 4.10(m, 1H), 4.07 (m, 1H), 3.38 (m, H), 3.30 (m, 1H), 3.09 (m, 3H), 2.95 (s,3H), 3.00 (s, 3H), 2.88 (s, 3H), 2.39 (s, 3H), 1.63 (m, 3H), 1.51 (m,3H), 1.44 (m, 1H), 1.39 (m, 1H), 1.16 (d, 3H), 1.11 (d, 3H).

¹³C NMR (CD₃)₂SO: δ=171.3, 170.8, 154.2, 150.2, 143.7, 134.1, 130.2,130, 127.6, 121.6, 68.2, 61.2, 53.5, 49, 36.3, 36.1, 35.7, 30.5, 23.8,21.4, 21.4, 21.

Example 109 Synthesis of N-(Benzylsulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-(4-N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 111 usingthe procedure described in Method 11.

Physical data was as follows:

MS (FAB) (M+H)⁺550.

Calcd. for: C₂₅H₃₁N₃O₇S₂; C, 54.62; H, 5.68; N 7.64.

Found: C 54.51; H 5.60; N 7.63.

Example 111 Synthesis of N-(Benzylsulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-(4-N,N-dimethylcarbamyloxy)phenylalanine tert-Butyl Ester

The title compound was prepared following the procedure outlined for thepreparation of Example 2 and by substituting the appropriate startingmaterials.

Physical data was as follows:

MS [M+H]⁺550.

Calcd. for: C₂₉H₃₉N₃O₇S₂; C, 57.52; H, 6.45; N, 6.94.

Found: C, 57.32; H, 6.52; N, 6.81.

Example 117 Synthesis ofN-(Methyl-pyrazole-4-sulfonyl)-L-prolyl-L-(4-N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

Substituting N-methyl-pyrazole sulfonyl chloride (see Dickson, U.S. Pat.No. 3,665,009 (May 23, 1972) and following the method for thepreparation of Example 56, gave the title compound.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.83 (s, 1H), 7.76 (s, 1H), 7.26 (m, 1H), 7.15 (m,2H), 7.00 (m, 2H), 4.69 (m, 1H), 3.95 (m, 1H), 3.93 (s, 3H), 3.38 (m,1H), 3.23-3.11 (m, 1H), 3.10-2.99 (m, 4H), 2.99 (s, 3H), 2.05 (m, 1H),1.66-1.46 (m, 3H), 1.44 (s, 9H).

¹³C NMR (CDCl₃): δ 170.7,169.9,154.9, 150.6,138.9, 133.2,132.5, 130.2,121.7, 117.9, 82.6, 62.4, 53.7, 49.7, 39.6, 37.7, 36.6, 36.4, 29.9,27.9, 24.2.

Example 132 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-prolyl-L4-(1,1-dioxothiomorphotin4-ylcarbonyloxy)-phenylalanine tert-Butyl Ester

Substituting thiamorpholine for N-methylpiperazine, and following themethod for the preparation of Example 4 and 14, gave the title compound.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.87-7.82 (m, 2H), 7.28-7.17 (m, 5H), 7.01 (d, 2H),4.71-4.69 (m, lH), 4.14-4.05 (m, 5H), 3.39-3.36 (m, 1H), 3.23-3.01 (m,7H), 2.05-2.03 (m, 1H), 1.58- 1.44 (m, 12H).

¹³C NMR (CDCl₃): δ=170.4,169.8, 153.0, 149.7,134.2, 130.6, 130.5, 121.3,116.8, 116.5, 82.6, 62.1, 53.6, 51.8, 49.5, 43.1, 42.7, 37.2, 29.7,27.8, 24.2.

Example 137 Synthesis ofN-(Methyl-pyrazole-4-sulfonyl)-L-prolyl-L-(4-N,N-dimethylcarbamyloxy)-phenylalanine Isopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 117.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.83 (s, 1H), 7.76 (s, 1H), 7.27 (d, 1H), 7.13 (d,2H), 7.01 (d, 2H) 5.06-5.02 (m, 1H), 4.80-4.73. (m, 1H), 3.97-3.94 (m,1H), 3.93 (s, 3H), 3.44-3.37 (m, 1H), 3.25-3.19 (m, 1H), 3.09-3.00 (m,5H), 2.97 (s, 3H), 2.06-2.02 (m, 1H), 1.66-1.48 (m, 3H), 1.23 (d, 6H).

¹³C NMR (CDCl₃): δ170.8, 170.5, 154.9, 150.6, 138.9, 132.9,32.5, 130.2,121.7, 117.8, 69.5, 62.3, 53.2, 49.7, 39.6, 37.1, 36.6, 36.3, 29.9,24.1, 21.6, 21.5. TABLE 10

R¹ R² R³ R⁵ R⁶ Ex. No. p-CH₃-φ- R²/R³ = cyclicp-[(4-methylpiperazin-1-yl)C(O)O-]benzyl- —O-n-butyl 99 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(4-methylpiperazin-1-yl)C(O)O-]benzyl- —O-cyclopentyl 100 3 carbonatoms (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O-]benzyl-—OCH(CH₃)₂ 101 3 carbon atoms (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(piperidin-4-yl)C(O)O-]benzyl- —OCH₂CH₃ 102 3 carbon atoms(L-pyrrolidinyl) φ-CH₂— R²/R³ = cyclicp-[(1-methylpiperidin-4-yl)C(O)O-]benzyl- —OCH₂CH₃ 103 3 carbon atoms(L-pyrrolidinyl) φ-CH₂— R²/R³ = cyclicp-[(4-methylpiperazin-1-yl)C(O)O-]benzyl- —OH 104 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(1-Boc-4-phenylpiperidin-4-yl)- —OCH₂CH₃ 105 3 carbon atomsC(O)O-]benzyl- (L-pyrrolidinyl) 1-methylimidazol- R²/R³ = cyclicp-[(CH₃)₂NC(O)O-]benzyl- —OC(CH₃)₃ 106 4-yl 3 carbon atoms(L-pyrrolidinyl) p-NH₂-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O-]benzyl-—OC(CH₃)₃ 107 3 carbon atoms (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(morpholin-4-yl)C(O)O-]benzyl- —OC(CH₃)₃ 108 3 carbon atoms(L-pyrrolidinyl) φ-CH₂— R²/R³ = cyclic p-[(CH₃)₂NC(O)O-]benzyl- —OH 109—CH₂—S—C(CH₃)₂— (L-5,5- dimethylthiazolidin- 4-yl) p-CH₃-φ- R²/R³ =cyclic p-[(CH₃)₂NC(O)O-]benzyl- —OH 110 —CH₂CH₂—NH—CH₂— (L-piperazinyl)φ-CH₂— R²/R³ = cyclic p-[(CH₃)₂NC(O)O-]benzyl- —OC(CH₃)₃ 111—CH₂—S—C(CH₃)₂— (L-5,5- dimethylthiazolidin- 4-yl) p-CH₃-φ- R²/R³ =cyclic p-[(CH₃)₂NC(O)O-]benzyl- —NH-adamantyl 112 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O-]benzyl-—NHCH₂C(O)OH 113 3 carbon atoms (L-pyrrolidinyl) p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NS(O)₂O-]benzyl- —OCH₃ 114 3 carbon atoms (L-pyrrolidinyl)p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O-]benzyl- —OC(CH₃)₃ 115—CH₂CH₂—NH—CH₂— (L-piperazinyl) p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O-]benzyl- —OC(CH₃)₃ 116 —CH₂CH₂— (Cbz)NHCH₂—[L-4-N-(Cbz)- piperazinyl] 1-methylpyrazol- R²/R³ = cyclicp-[(CH₃)₂NC(O)O-]benzyl- —OC(CH₃)₃ 117 4-yl 3 carbon atoms(L-pyrrolidinyl) 3-pyridyl R²/R³ = cyclic p-[(CH₃)₂NC(O)O-]benzyl- —OH118 3 carbon atoms (L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclicp-[(1-Boc-piperazin-4-yl)C(O)O-]benzyl- —OCH₂CH₃ 119 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclicp-[(morpholin-4-yl)C(O)O-]benzyl- —OCH₂CH₃ 120 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ —CH₃ H p-[(thiomorpholin-4-yl sulfone)-OC(CH₃)₃ 121 C(O)O-]benzyl- p-CH₃-φ —CH₃ Hp-[(thiomorpholin-4-yl)C(O)O-]benzyl- —OH 122 p-CH₃-φ R²/R³ = cyclicp-[(CH₃)₂NC(O)O-]benzyl- 2,4-dioxo- 123 3 carbon atoms tetrahydrofuran-(L-pyrrolidinyl) 3-yl (3,4-enol) p-CH₃-φ R²/R³ = cyclicp-[(piperazin-4-yl)C(O)O-]benzyl- —OH 124 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclicp-[(1-Boc-piperazin-4-yl)C(O)O-]benzyl- —OC(CH₃)₃ 125 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclicp-[(piperazin-4-yl)C(O)O-]benzyl- —OCH₂CH₃ 126 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclicp-[(4-acetylpiperazin-1-yl)C(O)O-]benzyl- —OCH₂CH₃ 127 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclicp-[(4-methanesulfonylpiperazin-1-yl)- —OCH₂CH₃ 128 3 carbon atomsC(O)O-]benzyl- (L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclic3-nitro-4-[(morpholin-4-yl)- —OH 129 3 carbon atoms C(O)O-]benzyl-(L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclicp-[(1-Boc-piperazin-4-yl)C(O)O-]benzyl- —OH 130 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ —CH₃ —C(CH₃)₃p-[(4-methylpiperazin-1-yl)C(O)O-]benzyl- —OC(CH₃)₃ 131 p-F-φ R²/R³ =cyclic p-[(1,1-dioxothiomorpholin-4-yl)- —OC(CH₃)₃ 132 3 carbon atomsC(O)O-]benzyl- (L-pyrrolidinyl) p-F-φ R²/R³ = cyclicp-[(morpholin-4-yl)C(O)O-]benzyl- —OC(CH₃)₃ 133 3 carbon atoms(L-pyrrolidinyl) p-CH₃-φ R²/R³ = cyclicp-[(morpholin-4-yl)C(O)O-]benzyl- —OH 134 —CH₂—CH₂—SO₂—CH₂— (L-1,1-dioxothiomorpholin- 3-yl) 1-methylpyrazol- R²/R³ = cyclicp-[(4-methylpiperazin-1-yl)C(O)O-]benzyl- —OC(CH₃)₃ 135 4-yl 3 carbonatoms (L-pyrrolidinyl) morpholin-4-yl R²/R³ = cyclicp-[(CH₃)₂NC(O)O-]benzyl- —OC(CH₃)₃ 136 3 carbon atoms (L-pyrrolidinyl)1-methylpyrazol- R²/R³ = cyclic p-[(CH₃)₂NC(O)O-]benzyl- —OCH(CH₃)₂ 1374-yl 3 carbon atoms (L-pyrrolidinyl)

Additional compounds prepared by the methods described above include thefollowing:

Example 138 Synthesis ofN-(1-Methylpyrazole4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L4-(N,N-dimethylcarbamyloxy)phenylalanine tert-Butyl Ester

The N-methylpyrazole sulfonyl chloride was prepared by addingN-methylpyrazole to chilled (0° C.) chlorosulfonic acid. The reactionmixture was allowed to warm to room temperature and then heated to 100°C. overnight under a stream of N₂. The reaction mixture was then cooledto room temperature and chilled to 0° C. To this solution was addedthionyl chloride (2.5 eq.) and the reaction was stirred at roomtemperature for 30 min., then warmed to 70° C. for two hours. Thereaction was cooled to room temperature and then chilled in an ice bath.Water and ice were slowly added to the reaction mixture to precipitate awhite solid which was collected by filtration. The desired sulfonylchloride was washed with cold water and hexane.

The title compound was then prepared following the procedure outlinedfor the preparation of Example 2 by substitution of the appropriatestarting materials, mp: 169- 170° C.

Example 139 Synthesis ofN-(1-Methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 138 usingthe procedure described in Method 11.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.94 (s, 1H); 7.79 (s, 1H); 7.25 (d, 2H, J=8.8Hz); 7.0(d, 2H, J=8.8 Hz); 5.15 (br s, 1H); 4.80 (m, 1H); 4.54 (d, 1H, J=9.1Hz); 4.39 (d, 1H, J=9.3 Hz); 3.93 (s, 3H); 3.88 (s, 1H); 3.23-3.02 (m,2H0; 3.07 (s, 3H); 2.98 (s, 3H); 1.27 (s, 3H); 1.14 (s, 3H).

¹³C NMR (CDCl₃): 173.86, 169.05, 155.23, 150.47, 139.21, 133.59, 133.15,130.53, 121.84, 117.57, 73,58, 54,71, 53.75, 50.42, 39.60, 37.18, 36.60,36.36, 35.11, 28.97, 23.95.

Example 140 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L4-(N-(1,4-dioxa-8-aza-spiro[4.5-decan-8-yl)carbonyloxy)phenylalanineEthyl Ester

The title compound was prepared following the procedure outlined forExample 4 by substitution of the appropriate starting materials.

Physical data was as follows:

MS (+ESI): 630 [M+H]⁺,

Anal. Calcd. for C₃H₃₉N₃O₉S.0.2CH₂Cl₂: C, 57.94; H, 6.14; N, 6.50.

Found: C, 57.73; H, 5.90; N, 6.47.

Example 141 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(N-(1,4-dioxa-8-aza-spiro [4.5]decan-8-yl)carbonyloxy)phenylalanine

The product of Example 140 was hydrolyzed using the procedure describedin Method 5 but employing methanol as the solvent and running thereaction at 25° C. for 24 h. The solvent was then evaporated, theresidue taken up in H₂O, washed with methylene chloride and lyophilizedto afford the title compound.

Physical data was as follows:

MS (+ESI): 619 [M+H]⁺.

Anal. Calcd. for C₂₉H₃₅N₃O₉SLi.1.5 H₂0: C, 53.37; H, 6.02; N, 6.44.

Found: C, 53.40; H, 5.58; N, 6.48.

Example 142 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L4-(4′-acetylpiperazin-1-ylcarbonyloxy)phenylalanine

The product of Example 127 was hydrolyzed using the procedure describedin Method 5 but employing methanol as the solvent and running thereaction at 25° C. for 24h. The solvent was then evaporated, the residuetaken up in H₂O, washed with methylene chloride and lyophilized toafford the title compound.

Physical data was as follows:

MS (+ESI): 587 [M+H]⁺.

Anal. Calcd. for C₂₈H₃₃N₄O₈SLi.3H₂0: C, 52.01; H, 6.08; N, 8.66.

Found: C, 52.03; H, 5.36; N, 8.04.

Example 143 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(4′-methanesulfonylpiperazin-1-ylcarbonyloxy)phenylalanine

The product of Example 128 was hydrolyzed using the procedure describedin Example 142.

Physical data was as follows:

MS (+ESI): 623 [M+H]⁺.

Anal. Calcd. for C₂₇H₃₃N₄O₉S₂Li.2 H₂0: C, 48.79; H, 5.61; N, 8.43.

Found: C, 48.66; H, 5.14; N, 8.04.

Example 144 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(4′-phenylpiperazin-1-ylcarbonyloxy)phenylalanine

The ethyl ester of the title compound was prepared following theprocedure outlined for Example 4 by substitution of the appropriatestarting materials. The ethyl ester was then hydrolyzed using theprocedure described in Example 142.

Physical data was as follows:

MS (-ESI): 619 [M−H].

Anal. Calcd. for C₃₂H₃₆N₄O₇SLi.2H₂0: C, 58.00; H, 5.93; N, 8.45.

Found: C, 57.65; H, 5.49; N, 8.13.

Example 145 Synthesis ofN-(Toluene4-sulfonyl)-L-prolyl-L4-(piperazin-1-ylcarbonyloxy)phenylalaninetert- Butyl Ester

The product of Example 125 (0.7 g, 1 mmol) was dissolved in methylenechloride (9 mL). The solution was cooled to 0° C. and trifluoroaceticacid (1.0 mL) was added and the resulting clear solution was stirred for4 h. The reaction solution was then diluted with additional methylenechloride (50 mL), washed with saturated sodium bicarbonate solution(3×50 mL), dried (K₂CO₃) and the solvent stripped off to give a whitesolid (0.465 g). Flash chromotography (9:1 CH₂Cl₂:EtOH) of this materialgave a clear oil which was washed several times with hexane to give awhite solid (0.289 g, 48%).

Physical data was as follows:

MS (+ESI): 601.7 [M+1]⁺.

Anal. Calcd. for C₃₀H₄₀N₄O₇S.0.25 CH₂CI₂: C, 58.42; H, 6.56; N, 9.01.

Found: C, 58.79; H, 6.51; N, 8.74.

Example 146 Synthesis of2-(Saccharin-2-yl)propionyl-L-4-(4′-methylpiperazin-1-ylcarbonyloxy)phenylalanine

The title compound was prepared from the product of Example 46 using theprocedure described in Method 11, mp 117-122° C. (with foaming).

Physical data was as follows:

Anal. Calcd. for C₂₅H₂₈N₄O₈S.1.5 H₂0: C, 52.53; H, 5.47; N, 9.80.

Found: C, 52.26; H, 5.36; N, 9.23.

Example 147 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(4′-methanesulfonyl piperazin-1-ylcarbonyloxy)phenylalanine tert-Butyl Ester

The title compound was prepared following the procedure outlined forExample 128 by substitution of the appropriate starting materials.

Physical data was as follows:

MS (+ESI): 696 [M+NH₄]⁺.

Anal. Calcd. for C₃₁H₄₂N₄O₉S_(2.)0.5 CH₂CI₂: C, 51.62; H, 6.00; N, 7.76.

Found: C, 51.55; H, 6.21; N, 7.60.

Example 148 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylanine(-tert- butoxycarbonyl-2-amino-2-methylpropyl) Ester

(BOC)₂O (96 mg, 0.44 mmol) was added to a solution of the product fromExample 9 (200 mg, 0.4 mmol.), N-Boc-2-amino-2-methyl-1-propanol (965mg, 0.5 mmol) and a catalytic amount of DMAP in THF (92 mL) containingpyridine (50 μl). The mixture was stirred at room temperature underargon for 48 h. The mixture was poured into 1N HCl and extracted withethyl acetate. The organic phase was washed (1N HCl), dried (MgSO₄) andthe solvent was removed under reduced pressure. The residue was purifiedby flash chromatography (EtOAc:hexanes 2:1) to give the desired compoundas an amorphous white foam (150 mg., 55%).

Physical data was as follows:

MS: [M+H]⁺ at 675.

MS (+ESI): [M+NH_(4]) ⁺ at 692 (100%).

Example 149 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine2- (Morpholin4-yl)ethyl Ester

The title compound was prepared following the procedure outlined forExample 148 by substituting 2-morpholinoethanol forN-Boc-2-amino-2-methyl- 1 -propanol.

Physical data was as follows:

Anal. Calcd. for C₃₀H₄₀N₄O₈S.0.5 H₂0: C, 57.58; H, 6.60; N, 8.95.

Found: C, 57.26; H, 6.29; N, 8.82.

Example 150 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(4-acetylpiperazin-1-ylcarbonyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure outlined forExample 127 by substitution of the appropriate starting materials.

Physical data was as follows: MS (+ESI): 660.4 [M+NH₄]+.

Anal. Calcd. for C₃₂N₄₂N₄0₈S *0.15 CH₂CI₂: C, 58.91; H, 6.50; N, 8.55.

Found: C, 58.64; H, 6.36; N, 8.40.

Example 151 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(4′-hydroxypiperidin-1-ylcarbonyloxy)phenylalanine tert-Butyl Ester

The title compound was prepared following the procedure outlined forExample 4 by substituting 4-piperidinol for N-methyl piperazine.

Physical data was as follows:

Anal. Calcd. for C₃H₄N₃0₈S -0.6 H₂O*0.22 EtOAc: C, 59.28; H, 6.86; N,6.51 Found: C, 58.92; H, 6.37; N, 6.47.

Example 152 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(N-(2′-(morpholin-4′-yl)ethyl)carbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure outlined forExample 4 by substituting 4-(2-aminoethyl)morpholine for N-methylpiperazine.

Physical data was as follows:

Anal. Calcd. for C₃₂H44N₄O₈S.0.25 H₂0: C,59.20; H, 6.91; N, 8.63

Found: C, 59.01; H, 6.54; N, 8.38.

Example 153 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(N-(1,4-dioxa-8-aza-spiro[4.5decan-8-yl)carbonyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure outlined forExample 4 by substitution of the appropriate starting materials.

Physical data was as follows:

MS (-ESI): 656 [M−H]⁻.

Anal. Calcd. for C₃₃H₄₃N₃O₉S.0.1 CH₂CI₂: C, 59.67; H, 6.54; N, 6.31.

Found: C, 59.83; H, 6.63; N, 6.66.

Example 154 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(N-(2′-hydroxyethyl)-N-methylcarbamyloxy)phenylalanine tert-Butyl Ester

The title compound was prepared folllowing the procedure outlined forExample 4 by substituting 2-(methylamino)ethanol for N-methylpiperazine.

Physical data was as follows:

Anal. Calcd. for C₂₉H₃₉N₃O₈S.0.5 H₂0: C, 58.18; H, 6.73; N, 7.02.

Found: C, 57.95; H, 6.5; N, 6.9.

Example 155 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(4′-formyloxypiperidin-1-ylcarbonyloxy)phenylalanine.

The title compound was prepared by treating the product of Example 151with formic acid overnight with stirring. The title compound wasobtained as a white foam (130 mg., 94%), following removal of excessformic acid.

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHZ) δ 12.8 (s, 1H); 8.23 (s, 1H); 8.09 (d, 1H);7.69 (d, 2H), 7.4 (d, 2H); 7.23 (d, 2H), 7.02 (d, 2H); 5.00 (m, 1H);4.45 (m, 1H); 4.10 (m, 1H); 3.6-3.8 (br, 2H); 3.4 (br s, 1H); 3.25 (m,2H); 3.10 (m, 2H); 2.95 (m, 1H); 2.35 (s, 3H); 1.95 (m, 2H); 1.56-1.75(m, 5H); 1.4 (m, 1H).

IR (KBr,cm⁻¹) 3400, 2950, 1720, 1680, 1510, 1430, 1325, 1250, 1150,1010, 650, 75, 540.

MS ((+)ESI, m/z (%)) 605 (100 [M+NH,1+).

Anal. Calcd. for C₂₈H₃₃N₃O₉S.0.66H₂0: C, 56.09; H, 5.77; N, 7.01.

Found: C, 56.14; H, 5.83; N, 6.78.

Example 156 Synthesis ofN-(Toluene4-sulfonyl)-L-prolyl-L-4-(4-hydroxypiperidin-1-ylcarbonyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure outlined forExample 4 by substitution of the appropriate starting materials, mp.64-67° C. (with foaming).

Physical data was as follows:

Anal. Calcd. for C₃₀H₃₉N₃0₈S *0.75 H₂O -0.1 EtOAc : C, 58.51; H, 6.67;N, 6.73.

Found: C, 58.55; H, 6.09; N, 6.78.

Example 157 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(4′-(2-hydroxyethyl)piperazin-1-ylcarbonyloxy)phenylalanine tert-Butyl Ester

The carbonate was prepared by treatment of Tos-Pro-Tyr-t-butyl esterwith 4-nitrophenyl chloroformate, followed by addition of N-(2-hydroxylethyl)piperazine (triethylamine, methylene chloride, chilled to 0° C.,then stirred at room temperature overnight). The crude product waspurified by flash chromatography (silica, 95:5 EtOAc:EtOH) to afford awhite solid, mp.158-160° C. (0.387 g, 58%).

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHZ) δ 8.15 (d, 1H, J=7.90 Hz); 7.70(d, 2H, J=6.59Hz); 7.40 (d, 2H, J=7.90 Hz); 7.23(d, 2H, J=8.56 Hz); 7.00 (d, 2H,J=8.56 Hz); 4.42 (m, 1H); 4.38 (m, 1H); 4.08 (m, 1H); 3.51 (m, 4H); 3.34(m, 3H); 3.09 (m, 1H); 2.99 (m, 2H); 2.43 (m, 6H); 2.39 (s, 3H); 1.59(m, 3H); 1.39 (m, 1H); 1.35 (s, 9H).

IR (KBr, cm⁻¹) 3505, 3400, 2990, 2930, 2890, 1730, 1700, 1670, 1510,1430, 1350, 1220, 1200, 1160, 670, 590, 545.

MS ((−)ESI, m/z (%)) 643 (98 [M−NH₄]).

Anal. Calcd. for C₃₂H44N₄O₈S: C, 59.61; H, 6.88; N, 8.69.

Found: C, 59.06; H, 6.95; N, 8.43.

Example 158 Synthesis of N-(Toluene-4-sulfonyl)-L-prolyl-L4-(N-(2¹-formyloxyethyl)-N- methylcarbamyloxy)phenylalanine

The title compound was prepared by treating the product of Example 154with formic acid overnight with stirring. The title compound wasobtained as a white foam (110 mg., 77%), following removal of excessformic acid.

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHz) δ 12.8 (s, 1H); 8.25 (d, 1H); 8.08 (d, 1H);7.69 (d, 2H), 7.40 (d, 2H); 7.22 (d, 2H), 6.98 (dd, 2H); 4.47 (m, 1H);4.35 (m, 1H); 4.27 (m, 1H); 4.10 (m, 1H); 3.65 (m, 1H); 3.55 (m, 1H);2.85-3.15 (overlapping m, 7H); 2.40 (s, 3H); 1.55 (m, 3H); 1.40 (m, 1H).

IR (KBr, cm⁻¹) 3420, 2910, 1725, 1510, 1400, 1340, 1270, 1150, 675, 590,550.

MS ((+)ESI, m/z (%)) 579 (100 [M+NH,1+).

Anal. Calcd. for C₂₆H₃₁N₃O₉S.0.66H₂0: C, 54.45; H, 5.68; N, 7.33

Found: C, 54.41; H, 5.60; N, 7.24.

Example 159 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(N-(2′-hydroxyethyl)-N-methylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure outlined forExample 4 by substitution of the appropriate starting materials, mp.49-52° C.

Physical data was as follows:

Anal Calcd. for C₂₈H₃₇N₃O₈S.0.5 H₂0: C, 57.52; H, 6.55; N, 7.19.

Found: C, 57.56; H, 6.38; N, 7.14.

Example 160 Synthesis ofN-(Toluene4-sulfonyl)-L-prolyl4-(N-(methoxycarbonylmethyl)carbamyloxy)phenylalaninetert-Butyl Ester

The carbonate was prepared by treatment of Tos-Pro-Tyr-t-butyl esterwith 4-nitrophenyl chloroformate, followed by addition of glycine methylester (triethylamine, methylene chloride, chilled to 0°, then stirred atroom temperature overnight). The crude product was purified by flashchromatography (silica, 3:2 EtOAc:hexane) to afford a white foam (0.640g, 35%).

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHz) δ8.15 (d, 1H, J=8.12 Hz); 8.12 (d, 2H, J=6.15Hz); 7.73 (d, 2H, J=8.34 Hz); 7.40 (d, 2H, J=7.90 Hz); 7.24 (d, 2H,J=8.56 Hz); 6.98 (d, 2H, J=8.34 Hz); 4.25 (m, 1H); 4.07 (m, 1H); 3.83(d, 2H, J=6.15 Hz); 3.64 (s, 3H); 3.32 (m, 1H); 3.02 (m, 3H); 2.39 (s,3H); 1.56 (m, 3H); 1.41 (m, 1H); 1.35 (s, 9H).

IR (KBr, cm⁻¹) 3400, 2990, 1745, 1680, 1500, 1370, 1350, 1200, 1160,670, 600.

MS ((+)ESI, m/z (%)) 621 (100[M+NH₄]⁺).

Anal. Calcd. for C₂₉H₃₇N₃O₉S: c, 57.70; H, 6.18; N, 6.96.

Found: C, 57.63; H, 6.11; N, 6.74.

Example 161 Synthesis ofN-(1-Methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl) thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine Isopropyl Ester

The title compound was prepared following the procedure outlined forExample 138 by substitution of the appropriate starting materials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.91 (s, 1H), 7.81 (s, 1H), 7.21 (d, 2H, J=8.2 Hz),7.03 (m, 3H); 5.03 (m, 1H), 4.84 (m, 1H), 4.55 (d, 1H), 4.42 (d, 1H),3.96 (s, 3H), 3.83 (s, 1H), 3.18-3.01 (m, 2H), 3.10 (s, 3H), 3.01 (s,3H), 1.28 (s, 3H), 1.24 (m, 6H), 1.17 (s, 3H).

¹³C NMR (CDCl₃): δ170.43, 166.31, 154.92, 150.68, 132.91, 132.88,130.34, 121.78, 117.69, 73.76, 69.61, 54,79, 53.2, 50.52, 39.61, 37.62,36.58, 36.35, 28.96, 24.02, 21.57, 21.49.

Example 162 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(4′-methoxypiperidin-1-ylcarbonyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure outlined forExample 156 by substitution of the appropriate starting materials. NMRdata was as follows:

¹H NMR (DSMO-d₆, 400 MHz) 8.10 (d, 1H); 7.72 (d, 2H); 7.41(d, 2H); 7.24(d, 2H); 7.02 (d, 2H); 4.92 (m, 1H); 4.45 (m, 1H); 4.10 (m, 1H); 3.8 (brs, 1H); 3.65 (br s, 1H); 3.40 (M, 2H); 3.25 (s, 3H); 2.95-3.15(overlapping m, 5H); 2.40 (s, 3H); 1.85 (br, 2H); 1.4-1.6 (m, 6H); 1.18(d, 3H); 1.12 (d, 3H).

IR(KBr, cm⁻¹) 3400, 2950, 1720, 1520, 1425, 1340, 1210, 1160, 1100, 625,590, 540.

MS ((+) ESI, m/z (%)) 633 [M+NH]⁺).

Example 163 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(4′-methoxypiperidin-1-ylcarbonyloxy)phenylalanine

The title compound was prepared from the product of Example 162 usingthe procedure described in Method 5.

NMR data was as follows:

¹H NMR (DMSO-d₆, 40(Y MHz) δ 12.8 (s, 1H); 8.10 (d, 1H); 7.72 (d, 2H);7.41 (d, 2H); 7.24 (d, 2H); 7.02 (d, 2H); 4.45 (m, 1H); 4.10 (m, 1H);3.8 (br s, 1H); 3.65 (br s, 1H); 3.40 (m, 2H); 3.25 (s, 3H); 2.95-3.15(overlapping m, 1H) 2.40 (s, 3H) 4.85 (br, 2H); 1.4-1.6 (m, 6H).

IR (KBr, cm⁻¹) 3400, 2950, 1720, 1520, 1425, 1340, 1210, 1160, 1100,625, 590, 540.

MS ((−)ESI, m/z (%)) 572 (100 [M−H]⁻).

Anal. Calcd. for C₂₈H₃₅N₃O₈S.0.33EtbAc.1H₂O: C, 56.73; H, 6.44; N, 6.77.

Found: C, 56.96; H. 6.01; N, 6.76.

Example 164 Synthesis ofN-(Toluene-4-sulfonyl)-L-4-oxoprolyl-L-4-(NN-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

Dichloromethane (7 mL) was cooled to −60° C. (chloroform/dry ice bath).Oxalyl chloride (0.15 mL) was added. The product from Example 165 (870mg) and dry DMSO (0.26 mL) were dissolved in dichloromethane (8 mL) andadded slowly to the above solution. The reaction was stirred at −60° C.for 30 minutes under dry conditions. Triethylamine (1.05 mL) was added.After 5 minutes, the dry ice bath was removed. The reaction was stirredat room temperature for 1 hour. The solvent was evaporated in vacuo.Ethyl acetate (30 mL) was added to the residue. The mixture was washedwith citric acid solution (5%, 2×30 mL) and saturated NaHCO₃ solution(2×30 mL); and finally with brine. The solution was dried over MgSO₄.The solvent was evaporated in vacuo, and the residue was flushed on asilica gel column to give 440 mg of the desired product, mp: 78-80° C.

Example 165 Synthesis ofN-(Toluene-4-sulfonyl)-L-trans-4-hydroxyprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

N-(Toluene-4-sulfonyl)-L-trans-4-hydroxyprolyl-L-4-(hydroxy)phenylalaninetert- butyl ester (1.60 g) and dimethylcarbamyl chloride (0.30 mL) weredissolved in DMF at 0C in an ice bath. Potassium carbonate powder (2.03g) was added to the solution. The ice bath was removed after 5 minutes.The reaction was stirred at room temperature for δhours. The solid wasfiltered. Ethyl acetate (40 mL) was added to the solution. The solutionwas washed with citric acid solution (5%, 40 mL) 2 times, and saturatedNaHCO₃ solution (40 mL) time. The solution was then washed with brineanddried with MgSO₄. The solvent was evaporated in vacuo to give 1.07 gof the title compound, mp: 170-172° C.

Example 166 Synthesis ofN-(3-Fluorobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

N-(3 -Fluorobenzenesulfonyl)-L-prolyl-L-4-(hydroxy)phenylalaninetert-butyl ester (700 mg) and dimethylcarbamyl chloride (0.2 mL) weredissolved in DMF (15 mL) at 0° C. in an ice bath. Potassium carbonatepowder (1.375 g) was added to the solution. The ice bath was removedafter 5 minutes. The reaction was stirred at room temperature forδhours. The solid was filtered. Ethyl acetate (20 mL) was added to thesolution. The solution was washed with citric acid solution (5%, 30 mL,2x), and saturated NaHCO₃ solution. The solution was then washed withbrine and dried with MgSO₄. The solvent was evaporated in vacuo to give890 mg of the title compound, mp: 107-109° C.

Example 167 Synthesis ofN-(Morpholino-sulfonyl)-L-prolyl-L-(4-N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

N-(Morpholino-sulfonyl)-L-proline was prepared using the proceduredescribed by Cheeseright et al., J. Chem. Soc. Perkin Trans. 1 1994, 12,1595-1600. The title compound was prepared following the proceduredescribed for the preparation of Example 2.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.13 (d, 2H), 7.03 (d, 2H), 6.92 (d, 1H), 4.71 (q,1H), 4.25 (t, 1H), 3.67 (t, 4H), 3.39 (dt, 1H), 3.28-3.19 (m, 1H), 3.23(t, 4H), 3.18 (dd, 1H), 3.08 (dd, 1H), 3.09 (s, 3H), 3.00 (s, 3H),2.16-2.08 (m, 2H), 1.98-1.86 (m, 1H), 1.78-1.66 (m, 1H), 1.45 (s, 9H).

¹³C NMR (CDCl₃): δ171.2, 170.4, 154.8, 150.7, 132.9, 130.3, 121.7, 82.7,66.3, 62.6, 53.3, 49.6, 46.2, 37.0, 36.6, 36.3, 30.5, 27.8, 24.7.

Example 168N-(Morpholino-sulfonyl)-L-prolyl-L-(4-N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 167 usingthe procedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ8.04 (d, 1H), 7.25 (d, 2H), 7.01 (d, 2H), 4.71-4.64 (m,1H), 4.22 (dd, 1H), 3.62 -3.50 (m, 4H), 3.43-3.31 (m, 2H), 3.24 (dd,1H), 3.11 (t, 4H), 3.09 (s, 3H), 3.03 (dd, 1H), 2.97 (s, 3H), 2.22-2.11(m, 1H), 1.98-1.80 (m, 3H).

¹³C NMR (CD₃OD): δ174.65, 174.58, 174.00, 156.60, 151.70, 135.30,131.20, 122.70, 67.10, 63.10, 54.59, 54.50, 50.6, 47.10, 37.10, 36.50,36.40, 32.0, 25.60.

Example 169 Synthesis ofN-(1-Methylpyrazole-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedures described forthe preparation of Examples 14 and 117.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.93 (s, 1H), 7.80 (s, 1H), 7.12 (d, 2H), 6.98 (d,2H), 6.44 (d, 10H), 4.95 (m, 1H), 4.66 (m, 1H), 4.04 (m, 2H), 3.98 (s,3H), 3.19 (m, 2H), 3.06 (m, 6H), 2.98 (m, 4H), 1.42 (m, 9H).

¹³C NMR (CDCl₃): δ170.58, 164.75, 154.91, 150.75, 139.33, 132.73,132.43, 130.43, 122.18, 119.66, 83.07, 56.02, 53.23, 50.03, 49.03,41.49, 39.63, 36.56, 36.31, 36.16, 27.87.

Example 170 Synthesis ofN-(2-Fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 90 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ 7.90 (m, 1H), 7.72 (m, 1H), 7.56 (d, 1H), 7.37 (m,2H), 7.20 (d, 2H), 7.07 (d,-2H), 5.18 (m 1H), 4.59-(m, -1H), 4.26 (m,1-H), 3.76 (m,-2H), 3.36 (m,-1H), 3.21 (m, 2H), 3.08 (m, 6H), 2.96 (s,3H).

¹³C NMR (CD₃OD): δ173.85, 168.04, 162.06, 158.69, 156.92, 152.06,137.69, 135.05, 131.83, 131.59, 129.77, 128.44, 128.26, 126.21, 123.17,119.04, 118.75, 57.04, 54.99, 52.08, 51.66, 43.36, 37.24, 36.83, 36.66.

Example 171 Synthesis ofN-(2,4-Difluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 92 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ8.88 (m, 1/2H), 8.14 (m, 1/2H), 7.90 (m, 1H), 7.64 (m,1H), 7.20 (m, 2H), 7.10 (m, 1H), 7.03 (m, 2H), 5.16 (m, 1H), 4.63 (m,1H), 4.28 (m, 1H), 3.75 (m, 2H), 3.41 (m, 1H), 3.15 (m, 5H), 3.02 (m,4H).

¹³C NMR (CD₃OD): δ173.91, 168.04, 156.93, 152.05, 135.15, 133.81,133.67, 131.60, 123.13, 113.48, 113.18, 107.38 ,107.02, 57.02, 55.02,52.29, 51.84, 43.45, 37.34, 36.83, 36.66.

Example 172 Synthesis ofN-(Toluene-4-sulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 49 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.67 (d, 2H), 7.32 (d, 2H), 7.21 (d, 2H), 7.10 (d,1H), 7.00 (d, 2H), 5.40 (bs, >1 H), 4.85 (m, 2H), 3.95 (m, 1H), 3.41 (m,1H), 3.07 (m, 6H), 2.98 (m, 4H), 2.62 (m, 1H), 2.41 (m, 5H), 2.13 (m,1H).

¹¹C NMR (CDCl₃): δ173.40, 168.49, 155.26, 144.44, 136.88, 132.95,1330.5, 130.30, 127.28, 122.08, 55.34, 53.45, 43.43, 36.62, 36.38,35.85, 25.25, 24.54, 21.43.

Example 173 Synthesis ofN-(Pyridine-3-sulfonyl)-L-(5,5-dimethyl-thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine Isopropyl Ester

The title compound was prepared following the procedure outlined for thepreparation of Example 56 by substitution of the appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 9.13 (m, 1H), 8.90 (m, 1H), 8.19 (m, 1H), 7.56 (m,1H), 7.23 (d, 2H), 7.04 (d, 2H), 6.93 (d, 1H), 5.07 (m, 1H), 4.85 (m,1H), 4.62 (d, 1H), 4.48 (d, 1H), 3.92 30 (s, 1H), 3.20-3.05 (m, 2H),3.12 (s, 3H), 3.03 (s, 3H), 1.32 -1.16 (m, 12H).

¹³C NMR (CDCl₃): δ 170.30, 167.75, 154.19, 150.67, 148.59, 135.72,132.94, 132.72, 130.27, 123.91, 121.78, 73.62, 69.64, 54.69, 53.12,50.48, 37.50, 36.53, 36.29, 29.05, 23.73, 21.54, 21.46.

Example 174 Synthesis ofN-(3-Fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 91 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ 7.68 (m, 3H), 7.44 (m, 1H), 7.20 (m, 2H), 7.01 (m,2H), 5.21 (m, 1H), 4.60 (m, 1H), 4.20 (m, 1H), 3.75 (m, 1H), 3.43 (m,1H), 3.21 (m, 3H), 3.02 (m, 4H), 2.96 (m, 4H).

¹³C NMR (CD₃OD): δ 173.98, 167.98, 165.89, 162.56, 156.94, 152.06,142.70 ,142.61, 135.11, 133.30, 133.19, 131.57, 124.71, 123.25, 122.21,121.93, 116.05, 115.71, 57.27, 54.87, 54.79, 51.29, 51.06, 43.24, 37.11,36.83.

Example 175 Synthesis ofN-(1-Methylpyrazole-4-sulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 169 usingthe procedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ 8.11 (s, 1H), 7.83 (s, 1H), 7.36 (d, 1H), 7.24 (d,2H), 7.02 (d, 2H),. 5.16 (m, 1H), 4.69 (m, 1H), 4.19 (m, 1H), 3.90 (s,3H), 3.81 (m, 2H), 3.33 (m, 3H), 3.10 (s, 3H), 3.02 (m, 4H).

¹³C NMR (CD₃OD): δ 174.07, 168.11, 156.93, 152.08, 140.12, 135.05,134.90, 131.67, 123.28, 121.82, 57.33, 54.77, 50.83, 50.64, 42.94,39.80, 37.02, 36.84, 36.76.

Example 176 Synthesis ofN-(4-tert-Butylbenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 88 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ 7.70 (d, 2H), 7.53 (d, 2H), 7.04 (d, 2H), 6.87 (d,2H), 5.09 (m, 1H), 4.48 (m, 1H), 3.99 (m, 1H), 3.60 (m, 1H), 2.90 (m,5H), 2.80 (m, 5H), 1.15 (s, 9H). ′³C NMR (CD₃OD): δ 173.95, 168.09,159.33, 156.88, 152.09, 137.52, 135.03, 131.54, 128.68, 128.15, 123.32,57.27, 54.81, 50.75, 43.04, 36.97, 36.82, 36.65, 36.16, 31.35.

Example 177 Synthesis ofN-(Toluene-4-sulfonyl)-(3,3-dimethyl)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 97 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.77 (d, 1H), 7.75 (d, 1H), 7.42-7.33 (m, 3.5H), 7.27(d, 1H), 7.19 (d; 0.5H), 7.10 (d, 1H), 7.03 (d, 1H), 5.07-5.00 (m,0.5H), 4.94-4.87 (m, 0.5), 3.67 (d, 1H), 3.58-3.52 (m, 1H), 3.35-3.25(m, 1H), 3.19-3.08 (m, 2H), 3.11 (s, 3H), 3.02 (s, 3H), 2.45 (s, 1.5 H),2.43 (s, 1.5H), 1.70-1.57 (m, 1H), 1.34-1.27 (m, 1H), 0.94 (s, 1.5H),0.75 (s, 1.5H), 0.54 (s, 6H).

¹³C NMR (CDCl₃): δ 174.6, 174.4, 171.8, 171.4, 155.7,150.5, 150.4,144.5, 144.4, 133.5, 132.6, 130.9, 130.6, 130.0, 129.9, 128.0, 127.9,122.2, 122.0, 71.2, 70.9, 53.3, 52.2, 47.3, 47.1, 43.0, 42.7, 38.1,37.9, 36.6, 36.4, 27.0, 26.8, 23.3, 23.0.

Example 178 Synthesis ofN-(2,5-Dichlorothiophene-3-sulfonyl)-L-prolyl-L4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 86 using theprocedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ 8.10 (d, 1H), 7.25 (d, 2H), 7.20 (s, 1H), 7.0 (d, 2H),4.65 (m, 1H), 4.35 (m, 1H), 3.55-3.35 (m, 2H), 3.30-3.20 (m, 2H),3.15-3.00 (m, 4H), 2.95 (s, 3H), 2.05-1.80 (m, 2H), 1.80- 1.65 (m, 2H).

¹³C NMR (CD₃OD):δ 174.2, 173.9, 156.9, 151.9, 135.9, 135.5, 132.3,131.6, 128.9, 128.6, 122.9, 63.1, 54.8, 54.7, 50.3, 37.4,.36.8, 36.7,32.1, 25.5.

Example 179 Synthesis ofN-(4-Methoxybenzenesulfonyl)-L-prolyl-L4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 180 usingthe procedure described in Method 7.

NMR data was as follows:

¹H NMR (CD₃OD): δ 7.78 (d, 2H), 7.27 (d, 2H), 7.09 (d, 2H), 7.02 (d,2H), 4.71 - 4.67 (m, 1H), 4.10-4.06 (m, 1H), 3.88 (s, 3H), 3.41-3.31 (m,1H), 3.28-3.07 (m, 6H), 2.97 (s, 3H), 1.81-1.50 (m, 4H).

¹³C NMR (CD₃OD): δ 168.3, 168.2, 159.2, 150.9, 145.9, 129.5, 125.6,125.3, 123.5, 116.9, 109.6, 57.2, 50.2, 48.7, 44.6, 31.4, 30.8, 30.6,25.7, 19.3.

Example 180 Synthesis ofN-(4-Methoxybenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared using the procedure described in Example2 by substitution of the appropriate starting materials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.76 (d, 2H), 7.34 (d, 1H), 7.14 (d, 2H), 7.03-6.97(m, 4H), 5.08-5.04 (m, 1H), 4.77 (m, 1H), 4.05-4.03 (m, 1H), 3.86 (s,3H), 3.37-3.34 (m, 1H), 3.26-3.19 (m, 1H), 3.10-3.01 (m, 4H), 2.98 (s,3H), 2.02 (m, 1H), 1.56-1.46 (m, 3H), 1.25 (d, 6H).

¹³C NMR (CDCl₃): δ 170.8, 170.3, 163.4, 154.8, 150.5, 132.9, 130.1,129.9, 127.6, 121.6, 114.3, 69.4, 62.1, 55.4, 53.2, 49.5, 37.1, 36.5,36.2, 29.7, 24.0, 21.5, 21.4.

Example 181 Synthesis ofN-(Toluene-4-sulfonyl)-L-(1-oxo-thiomorphotin-3-carbonyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 182 usingthe procedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ 7.90 (m, 1H), 7.78 (m, 2H), 7.40 (m, 2H), 7.26 (m,2H), 7.03 (m, 2H), 5.14 (m, 1H), 4.64 (m, 2H), 3.81 (m, 1H), 3.71 (m,2H), 3.19 (m, 1H), 3.14 (m, 3H), 3.02 (m, 4H), 2.84 (m, 1H), 2.60 (m,1H), 2.42 (m, 4H), 2.21 (m, 1H).

¹³C NMR (CD₃OD): δ 174.22, 173.93, 169.59, 156.88, 152.08, 152.05,146.44, 146.26, 137.75, 137.63, 135.61, 134.96, 131.79, 131.64, 131.55,131.39, 128.75, 128.66, 123.35, 123.06, 57.03, 54.88, 54.66, 51.64,42.69, 42.51, 40.34, 37.12, 36.83, 36.66, 32.76, 21.51.

Example 182 Synthesis ofN-(Toluene-4-sulfonyl)-L-(1-oxo-thiomorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 49. The oxidation of the thiomorpholine groupto the 1-oxo-thiomorpholine group was per Larsson and Carlson (ActaChemica Scan. 1994, 48, 517-525).

NMR data was as follows:

¹H NMR (CDCl₃):δ 7.72 (m, 2H), 7.69 (m, 2H), 7.31 (m, 2H), 7.11 (m, 2H),7.07 (m, 2H), 6.96 (m, 2H), 4.79 (m, 1H), 4.54 (m, 1H), 3.80 (m, 4H),3.04 (4H), 2.92 (m, 3H), 2.64 (m, 1H), 2.43 (m, 4H), 1.44 (s, 3H), 1.36(s, 6H).

¹³C NMR (CDCl₃): δ 169.8, 166.5, 166.,3 154.6, 150.5, 150.4, 144.9,144.4, 135.7, 135.3, 132.8, 130.5, 130.1, 29.9, 127.4, 126.9, 122.1,121.4, 82.6, 82.2, 55.6, 53.9, 53.1, 50.6, 48.1, 47.8, 41.7, 40.5, 38.3,36.4, 36.1, 31.1, 27.5, 21.2.

Example 183 Synthesis ofN-(3,4-Difluorobenzenesulfonyl)-L-prolyl4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 by substitution of the appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.72-7.60 (m, 2H), 7.87-7.37 (m, 1H), 7.13-7.11 (m,3H), 7.01 (d, 2H), 5.08-5.04 (m, 1H), 4.81-4.74 (m, 1H), 4.09-4.06 (m,1H), 3.39-3.35 (m, 1H), 3.26-3.19 (m, 1H), 3.12-2.97 (m, 8H), 2.06-2.03(m, 1H), 1.66-1.57 (m, 3H), 1.26 (d, 6H).

¹³C NMR (CDCl₃): δ 170.50, 170.40, 154.90, 153.60, 150.70, 150.30,133.30, 132.90, 130.10, 125.00, 121.80, 121.80, 118.50, 112.80, 69.60,62.20, 53.20, 49.60, 37.10, 36.60, 36.30, 30.10, 24.20, 21.59, 21.56.

Example 184 Synthesis ofN-(3,4-Difluorobenzenesulfonyl)-L-prolyl-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 183 usingthe procedure described in Method 7.

NMR data was as follows:

¹H NMR (CD₃OD): δ 8.10 (d, 1H), 7.84-7.77 (m, 1H), 7.69-7.65 (m, 1H),7.53-7.45 (m, 1H), 7.28 (d, 2H), 7.02 (d, 2H), 4.72-4.68 (m, 1H),4.19-4.16 (m, 1H), 3.43-3.39 (m, 1H), 3.31-3.21 (m, 2H), 3.13-3.05 (m,4H), 2.97 (s, 3H), 1.86-1.61 (m, 4H).

¹³C NMR (CD₃D): δ 174.2, 174.1, 164.7, 156.9, 154.9, 152.0, 151.6,135.8, 135.6, 131.6, 129.7, 122.9, 119.7, 118.8, 63.1, 54.7, 50.5, 37.4,36.8, 36.6, 31.9, 25.5.

Example 185 Synthesis ofN-(3,4-Difluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared using the procedure described in Example92 by substitution of the appropriate starting materials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.71 (m, 2H), 7.33 (m, 1H), 7.07 (d, 2H), 6.91 (d,2H), 6.36 (d, 1H), 4.95 (m, 1H), 4.61 (m, 1H), 4.03 (m, 2H), 3.16 (m,2H), 3.13 (m, 4H), 3.07 (m, 1H), 2.93 (s, 9H), 1.43 (s, 9H).

¹³C NMR (CDCl₃): δ 170.07, 169.45, 164.42, 155.06, 155.44, 154.81,152.21, 152.17, 150.58, 148.81, 148.64, 134.90, 134.85, 132.41, 130.29,124.82, 124.71, 124.66, 121.97, 119.07, 118.76, 117.52, 117.23, 82.92,55.98, 53.20, 50.10, 49.40, 41.76, 36.41, 36.16, 35.99, 27.64.

Example 186 Synthesis ofN-(3,4-Difluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 185 usingthe procedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ 6.22 (m, 1H), 6.03 (m, 1H), 5.84 (m, 1H), 5.58 (m,2H), 5.38 (m, 2H), 3.33 (m, 1H), 3.01 (m, 1H), 2.57 (m, 1H), 2.14 (m,1H), 1.91 (m, 1H), 1.66 (m, 3H), 1.44 (s, 3H), 1.35 (m, 3H), 1.32 (s,3H).

¹³C NMR (CD₃OD): δ 173.97, 167.89, 156.94, 153.53, 152.07, 150.00,137.48, 135.17, 131.63, 126.54, 126.43, 123.20, 120.21, 119.96, 118.84,118.57, 57.25, 54.82, 51.29, 49.86, 43.29, 37.21, 36.85, 36.67.

Example 187 Synthesis ofN-(Toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-(thiomorpholin-4-ylcarbonyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared using the procedure described in Example82 by substitution of the appropriate starting materials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.64 (d, 2H), 7.33 (d, 2H), 7.25 (d, 2H), 7.08-6.97(m, 3H), 4.76 (m, 1H), 4.57 (d, 1H), 4.38 (d, 1H), 3.83 (s, 1H),3.95-3.78 (m, 4H), 3.09 (m, 2H), 2.69 (m, 4H), 2.43 (s, 3H), 1.44 (s,9H), 1.16 (s, 3H), 1.08 (s, 3H).

¹³C NMR (CDCl₃): δ 169.78, 168.36, 153.53, 150.28, 144.84, 133.53,132.76, 130.51, 130.03, 128.19, 121.58, 82.69, 73.42, 54.56,53.78,50.46, 47.05, 46.40, 37.80, 29.06, 27.76, 27.37, 27.04, 23.86, 21.52.

Example 188 Synthesis ofN-(Toluene4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-(thiomorpholin-4-ylcarbonyloxy)phenylalanine

The title compound was prepared from the product of Example 187 usingthe procedures described in Method 11.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.77 (d, 2H), 7.37 (d, 2H), 7.28 (d, 2H), 7.22 (d,1H), 7.03 (d, 2H), 5.35 (brs, 1H), 4.91 (m, 1H), 4.60 (d, 1H), 4.39 (d,1H), 3.91 (s, 1H), 3.96-3.28 (m, 4H), 3.30-3.07 (m, 2H), 2.67 (m, 4H),2.45 (s, 3H), 1.10 (s, 3H), 1.08 (s, 3H).

¹³C NMR (CDCl₃): δ 173.09, 169.45, 153.81, 150.28, 145.02, 133.42,132.61, 130.60, 130.12, 128.13, 121.86, 73.28, 54.51, 53.31, 50.48,47.08, 46.47, 36.97, 28.97, 27.35, 27.03, 23.70, 21.52.

Example 189 Synthesis ofN-(1-Methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineEthyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 117 by substitution of the appropriatestarting materials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.89 (s, 1H), 7.81 (s, 1H), 7.19 (d, 2H), 7.00 (m,3H), 4.87 (m, 1H), 4.54 (d, 1H), 4.42 (d, 1H), 4.18 (q, 2H), 3.95 (s,3H), 3.81 (s, 1H), 3.11 (m, 2H), 3.08 (s, 3H), 2.99 (s, 3H), 1.30 (s,3H), 1.25 (t, 3H), 1.16 (s, 3H).

¹³C NMR (CDCl₃): δ 170.98, 168.34, 154.91, 150.71, 139.62, 132.88,130.28, 121.85, 117.71, 73.77, 61.66, 54.80, 53.16, 50.53, 39.64, 37.63,36.60, 36.36, 28.98, 24.00, 13.92.

Example 190 Synthesis ofN-(Pyridine-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 191 usingthe procedure described in Method 11.

NMR data was as follows:

¹H NMR (CDCl₃): δ 9.09 (s, 1H), 8.82 (m, 1H), 8.20 (m, 1H), 7.56 (m,1H), 7.23 (d, 2H), 7.07 (d, 1H), 5.58 (brs, 1H), 4.83 (m, lh), 4.56 (m,2H), 4.07 (s, 1H), 3.14 (m, 2H), 3.07 (s, 3H), 2.99 (s, 3H), 1.26 (s,3H), 1.18 (s, 3H).

¹³C NMR (CDCl₃): δ 173.04, 168.29, 155.16, 153.39, 150.60, 147.96,136.43, 133.91, 133.06, 130.66, 130.50, 124.65, 122.14, 121.91, 73.43,54.58, 53.21, 50.38, 37.18, 36.64, 36.38, 29.25, 23.64.

Example 191 Synthesis ofN-(Pyridine-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 56 and substitution of appropriate startingmaterials.

Physical data was as follows:

MS: [M+H]⁺593

Anal. Calcd. for C₂₇H₃₆N₄O₇S₂.0.5 H₂0: C, 53.88; H, 6.07; N, 9.27.

Found: C, 53.98; H, 6.07; N, 9.27.

Example 192 Synthesis ofN-(Pyridine-2-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

Substituting 2-pyridinesulfonyl chloride (see Corey et al., J Org. Chem.1989, 54, 389-393) and following the method for the preparation ofExample 56, gave the title compound.

NMR data was as follows:

¹H NMR (CDCl₃): δ 8.59 (d, 1H), 8.00-7.89 (m, 2H), 7.78 (d, 1H),7.53-7.49 (m, 1H), 7.16 (d, 2H), 7.01 (d, 2H), 5.05-4.99 (m, 1H),4.85-4.78 (m, 1H), 4.60-4.57 (m, 1H), 3.44 -3.35 (m, 2H), 3.25-3.19 (m,1H), 3.07 (s, 3H), 3.06-3.01 (m, 1H), 2.97 (s, 3H), 2.19-2.13 (m, 1H),1.88-1.71 (m, 2H), 1.55 (m, 1H), 1.22-1.19 (m, 6H).

¹³C NMR (CDCl₃): δ 170.90, 170.30, 156.20, 154.80, 150.50, 150.00,138.00, 133.10, 130.10, 127.00, 123.40, 121.60, 69.20, 62.80, 53.30,49.60, 37.20, 36.40, 36.20, 29.80, 24.30, 21.42, 21.40.

Example 193 Synthesis ofN-(Pyridine-2-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 192 usingthe procedure described in Method 7.

NMR data was as follows.

¹H NMR (CD₃OD): δ 8.67 (d, 1H), 8.27 (d, 1H), 8.07-8.02 (m, 1H),7.96-7.91 (m, 1H), 7.65-7.61 (m, 1H), 7.27 (d, 2H), 7.01 (d, 2H),4.72-4.69 (m, 1H), 4.58-4.54 (m, 1H), 3.44-3.37 (m, 2H), 3.28-3.24 (m,1H), 3.13-3.05 (m, 4H), 2.96 (s, 3H), 1.94-1.89 (m, 2H), 1.70-1.63 (m,2H).

¹³C NMR (CD₃OD): δ 174.5, 174.4, 174.2, 157.7, 156.9, 151.9, 139.9,135.6, 131.6, 128.8, 124.7, 122.9, 64.1, 54.8, 54.7, 50.9, 37.5, 36.8,36.7, 31.9, 25.6.

Example 194 Synthesis ofN-(Pyridine-2-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 192 by substitution of the appropriatestarting materials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 8.64-8.62 (m, 1H), 7.98-7.92 (m, 2H), 7.56-7.51 (m,1H), 7.28-7.21 (m, 3H), 7.01 (d, 2H), 5.01-4.97 (m, 1H), 4.88-4.85 (m,2H), 4.80 (d, 1H), 4.63 (d, 1H), 4.19 (s, 1H), 3.11-3.07 (m, 5H), 2.98(s, 3H), 1.28 (s, 3H), 1.26-1.18 (m, 9H).

¹³C NMR (CDCl₃): δ 170.3, 168.4, 155.5, 154.9, 150.7, 150.4, 138.2,133.0, 130.4, 127.5, 123.5, 121.8, 73.5, 69.5, 54.7, 53.3, 51.0, 37.6,36.6, 36.4, 29.3, 23.9, 21.52, 21.50.

Example 195 Synthesis ofN-(Pyridine-2-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 194 usingthe procedure described in Method 7.

NMR data was as follows:

¹H NMR (CD₃OD): δ 8.70-8.69 (m, 1H), 8.07-8.01 (m, 1H), 7.92-7.89 (m,1H), 15 7.67-7.63 (m, 1H), 4.77-4.67 (m, 3H), 4.30 (s, 1H), 3.23-3.06(m, 5H), 2.97 (s, 3H), 1.27-1.18 (m, 6H).

¹³C NMR (CD₃OD): δ 174.1, 171.2, 157.0, 151.9, 151.6, 139.9, 135.7,131.8, 131.7, 129.0, 124.6, 122.9, 74.3, 61.6, 55.7, 54.9, 51.9,37.6,36.8, 36.7, 30.1, 24.9.

Example 196 Synthesis ofN-(Toluene-4-sulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 49 by substitution of the appropriatestarting materials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.67 (d, 2H), 7.30 (d, 2H), 7.12 (d, 2H), 6.97 (d,2H), 6.86 (d, 1H), 5.05 (m, 1H), 4.70 (m, 2H), 3.90 (m, 1H), 3.31 (m,1H), 3.06 (m, 4H), 2.97 (s, 3H), 2.68 (m, 1H), 2.50 (m, 1H), 2.44 (s,3H), 2.29 (m, 1H), 2.13 (m, 1H), 1.24 (s, 3H), 1.22 (s, 3H).

¹³C NMR (CDCl₃): δ 170.35, 167.55, 155.00, 150.61, 144.20, 136.80,132.51, 130.24, 130.14, 127.20, 121.82, 69.48, 55.14, 53.55, 43.26,36.43, 36.16, 25.21, 24.56, 21.48, 21.31.

Example 197 Synthesis ofN-(3-Fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 by substitution of the appropriate startingmaterial.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.68 (d, 1H), 7.61-7.52 (m, 2H), 7.36 (dt, 1H), 7.21(d, 2H), 7.02 (d, 2H), 6.94 (d, 1H), 5.05 (sept, 1H), 4.85 (q, 1H), 4.59(d, 1H), 4.41 (d, 1H), 3.88 (s, 1H), 3.17-3.03 (m, 2H), 3.09 (s, 3H),3.00 (s, 3H), 1.25 (d, 3H), 1.23 (d, 3H), 1.16 (s, 3H), 1.12 (s, 3H).

¹³C NMR (CDCl₃): δ 170.3, 168.1, 162.6, 154.9,150.7, 137.9, 132.8,131.3, 130.4, 123.9, 121.8, 121.0, 115.4, 73.5, 69.6, 54.5, 53.2, 50.5,37.6, 36.6, 36.3, 29.0, 23.7, 21.6, 21.5.

Example 198 Synthesis ofN-(2-Fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 by substitution of the appropriate startingmaterial.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.92-7.87 (m, 1H), 7.67-7.59 (m, 1H), 7.33-7.24 (m,2H), 7.21 (d, 2H), 7.03 (d, 2H), 6.93 (d, 1H), 5.03 (Sept, 1H), 4.83 (q,1H), 4.67 (d, 1H), 4.63 (d, 1H), 4.03 (s, 1H), 3.16-3.03 (m, 2H), 3.09(s, 3H), 3.00 (s, 3H), 1.31 (s, 3H), 1.24 (d, 3H), 1.22 (d, 3H), 1.19(s, 3H).

¹³C NMR (CDCl₃): δ 170.3, 168.1, 159.2, 154.9, 150.7, 136.0, 132.9,132.0, 130.3, 124.6, 121.8, 117.6, 73.3, 69.6, 54.8, 53.2, 50.3, 37.6,36.6, 36.3, 29.1, 23.9, 21.6, 21.5.

Example 199 Synthesis ofN-(3,4-Difluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 by substitution of the appropriate startingmaterial.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.77-7.71 (m, 1H), 7.70-7.65 (m, 1H), 7.40-7.31 (m,1H), 7.20 (d, 2H), 7.02 (d, 2H), 6.87 (d, 1H), 5.05 (Sept, 1H),4.88-4.82 (m, 1H), 4.55 (d, 1H), 4.44 (d, 1H), 3.91 (s, 1H), 3.17-3.03(m, 2H), 3.09 (s, 3H), 3.00 (s, 3H), 1.25 (d, 3H), 1.23 (d, 3H), 1.23(s, 3H), 1.18 (s, 3H).

¹³C NMR (CDCl₃): δ 170.4, 167.9, 154.9, 150.7, 133.1, 132.7, 130.4,124.4, 121.8, 118.5, 118.0, 73.6, 69.7, 54.6, 53.1, 50.5, 37.6, 36.6,36.3, 29.2, 23.7, 21.6, 21.5.

Example 200 Synthesis ofN-(3,5-Difluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyi-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 by substitution of the appropriate startingmaterial.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.77-7.71 (m, 1H), 7.70-7.65 (m, I H), 7.40-7.31 (m,1H), 7.20 (d, 2H), 7.02 (d, 2H), 6.87 (d, 1H), 5.05 (Sept, 1H),4.88-4.82 (m, 1H), 4.55 (d, 1H), 4.44 (d, 1H), 3.91 (s, 1H), 3.17-3.03(m, 2H), 3.09 (s, 3H), 3.00 (s, 3H), 1.25 (d, 3H), 1.23 (d, 3H), 1.23(s, 3H), 1.18 (s, 3H).

¹³C NMR (CDCl₃): δ 170.4, 167.9, 154.9, 150.7, 133.1, 132.7, 130.4,124.4, 121.8, 118.5, 118.0, 73.6, 69.7, 54.6, 53.1, 50.5, 37.6, 36.6,36.3, 29.2, 23.7, 21.6, 21.5.

Example 201 Synthesis ofN-(2,4-Difluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 by substitution of the appropriate startingmaterial.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.94-7.86 (m, 1H), 7.20 (d, 2H), 7.03 (d, 2H),7.02-6.95 (m, 2H), 6.88 (d, 1H), 5.03 (September, 1H), 4.82 (q, 1H),4.67 (d, 1H), 4.61 (d, 1H), 4.01 (s, 1H), 3.16-3.03 (m, 2H), 3.09 (s,3H), 3.00 (s, 3H), 1.36 (s, 3H), 1.23 (d, 3H), 1.21 (d, 3H), 1.20 (s,3H).

¹³C NMR (CDCl₃): δ 170.3, 167.9, 154.9, 150.7, 133.7 ,132.8, 130.3,121.8, 112.1, 106.1, 73.4, 69.6, 54.9, 53.2, 50.4, 37.6, 36.6, 36.3,29.1, 23.9, 21.6, 21.5.

Example 202 Synthesis ofN-(4-Chlorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyi-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 by substitution of the appropriate startingmaterial.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.82 (d, 2H), 7.53 (d, 2H), 7.21 (d, 2H), 7.02 (d,2H), 6.93 (d, 1H), 5.05 (Sept., 1H), 4.89-4.82 (m, 1H), 4.55 (d, 1H),4.41 (d, 1H), 3.87 (s, 1H), 3.17-3.03 (m, 2H), 3.09(s 3H), 3.00 (s, 3H),125 (d,3H), 123 (d,3H) 1.16 (s, 6H)

¹³C NMR (CDCl₃): δ 170.3, 168.1, 154.9, 150.7, 140.4, 134.5, 132.8,130.4, 129.7, 129.5, 121.8, 73.5, 69.6, 54.6, 53.1, 50.5, 37.6, 36.6,36.3, 29.1, 23.8, 21.6, 21.0.

Example 203 Synthesis ofN-(3-Chlorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 by substitution of the appropriate startingmaterial.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.88 (t, 1H), 7.78-7.75 (m, 1H), 7.64-7.61 (m, 1H),7.51 (t, 1H), 7.21 (d, 2H), 7.02 (d, 2H), 6.92 (d, 1H), 5.05 (sept, 1H),4.89-4.82 (m, 1H), 4.58 (d, 1H), 4.40 (d, 1H), 3.88 (s, 1H), 3.18-3.03(m, 2H), 3.09 (s, 3H), 3.00 (s, 3H), 1.25 (d, 3H), 1.23 (d, 3H), 1.16(s, 3H), 1.14 (s, 3H).

¹³C NMR (CDCl₃): δ 170.3, 168.0, 154.9, 150.7, 137.7, 135.7, 133.9,132.8, 130.7, 130.3, 127.9, 126.2, 121.8, 73.6, 69.96, 54.5, 53.2, 50.5,37.6, 36.6, 36.3, 29.1, 23.7, 21.6, 21.5.

Example 204 Synthesis ofN-(2-Chlorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 by substitution of the appropriate startingmaterial.

NMR data was as follows:

¹H NMR (CDCl₃): δ 8.08 (dd, 1H), 7.54-7.52 (m, 2H), 7.45-7.39 (m, 1H),7.19 (d, 2H), 7.02 (d, 2H), 6.79 (d, 1H), 5.00 (sept, 1H), 4.78 (d, 1H),4.75-4.68 (m, 1H), 4.69 (d, 1H), 4.19 (s, 1H), 3.09 (s, 3H), 3.06 (d,2H), 3.00 (s, 3H), 1.38 (s, 3H), 1.23 (s, 3H), 1.23 (d, 3H), 1.19 (d,3H).

¹³C NMR (CDCl₃): δ 170.3, 168.1, 154.9, 150.7, 135.6, 134.4, 132.8,132.7, 132.4, 130.3,127.3, 121.8, 73.3, 69.5, 54.7, 53.3, 50.4, 37.6,36.6, 36.3, 29.6, 23.7, 21.6, 21.5.

Example 205 Synthesis ofN-(3,4-Dichlorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 by substitution of the appropriate startingmaterial.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.97 (d, 1H), 7.70 (dd, 1H), 7.63 (d, 1H), 7.20 (d,2H), 7.02 (d, 2H), 6.86 (d, 1H), 5.05 (sept, 1H), 4.89-4.82 (m, 1H),4.55 (d, 1H), 4.43 (d, 1H), 3.92 (s, 1H), 3.17-3.03 (m, 2H), 3.09 (s,3H), 3.00 (s, 3H), 1.26 (d, 3H), 1.22 (d, 3H), 1.23 (s, 3H), 1.18 (s,3H).

¹³C NMR (CDCl₃): δ 170.3, 167.9, 154.9, 150.7, 138.7, 136.1, 134.2,132.7, 131.4, 130.3, 129.8, 127.1, 121.8, 73.6, 69.7, 54.6, 53.1, 50.5,37.5, 36.6, 36.3, 29.2, 23.7, 21.6, 21.5.

Example 206 Synthesis ofN-(3,5-Dichlorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 by substitution of the appropriate startingmaterial.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.76 (d, 2H), 7.62 (t, 1H), 7.20 (d, 2H), 7.03 (d,2H), 6.85 (d, 1H), 5.05 (sept, 1H), 4.89-4.82 (m, 1H), 4.57 (d, 1H),4.42 (d, 1H), 3.92 (s, 1H), 3.18-3.04 (m, 2H), 3.09 (s, 3H), 3.00 (s,3H), 1.25 (d, 3H), 1.23 (d, 3H), 1.27 (s, 3H), 1.18 (s, 3H).

¹³C NMR (CDCl₃): δ 170.3, 167.8, 154.9, 150.7, 139.1, 136.5, 133.7,132.7, 130.3, 126.2, 121.8, 73.7, 69.7, 54.6, 53.1, 50.5, 37.5, 36.6,36.3, 29.2, 23.7, 21.6, 21.5.

Example 207 Synthesis ofN-(3-Chlorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared using the procedure described in Example92 by substitution of the appropriate starting materials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.85 (m, 1H), 7.76 (m, 1H), 7.63 (m, 1H), 7.53 (m,1H), 7.06 (d, 2H), 6.96 (d, 2H), 6.37 (m, 1H), 5.01 (m, 1H), 4.62 (m,1H), 4.01 (m, 2H), 3.26 (m; 1H), 3.06 (s, 3H), 2.96 (m, 7H), 1.49 (s,9H).

¹³C NMR (CDCl₃): δ 170.0, 164.5, 154.9, 150.6, 140.0, 136.1, 134.2,132.5, 131.3, 130.2, 127.4, 125.5, 122.2, 82.8, 56.0, 53.3, 49.9, 49.2,41.7, 36.5, 36.3, 36.0, 27.8.

Example 208 Synthesis ofN-(3,4-Dichlorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared using the procedure described in Example92 by substitution of the appropriate starting materials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.91 (m, 1H), 7.66 (m, 2H), 7.06 (d, 2H), 6.94 (d,2H), 6.33 (m, 1H), 4.98 (m, 1H), 4.60 (m, 1H), 3.49 (m, 3H), 3.12 (m,2H), 3.04 (s, 3H), 3.00 (m, 2H), 2.94 (s, 3H), 1.44 (s, 9H).

¹³C NMR (CDCl₃): δ 170.0, 164.3, 154.8, 150.6, 138.8, 137.9, 134.3,132.4 132.0, 130.3, 129.2, 126.4, 122.1, 83.0, 55.5, 53.1, 50.2, 49.5,41.8, 36.5, 36.2, 36.0, 27.7.

Example 209 Synthesis ofN-(4-Methoxybenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 by substitution of the appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.81 (d, 2H), 7.22 (d, 2H), 7.06-6.99 (m, 5H), 5.04(sept, 1H), 82. (m, 1H), 4.56 (d, 1H), 4.39 (d), 3.88 (s, 3H), 3.83 (s,1H), 3.17-3.03 (m, 2H), 3.09 (s, 3H), 3.00 (s, 3H), 1.25 (d, 3H), 1.22(d, 3H), 1.15 (s, 3H), 1.12 (s, 3H).

¹³C NMR (CDCl₃): δ 170.3, 168.5, 163.8, 154.9,150.7, 132.9, 130.4,130.3, 127.4, 121.7, 114.5, 73.5, 69.5, 55.6, 54.6, 53.2, 50.5, 37.7,36.6, 36.3, 29.1, 23.9, 21.6, 21.5.

Example 210 Synthesis ofN-(3-Methoxybenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 by substitution of the appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.47-7.45 (m, 2H), 7.37-7.36 (m, 1H), 7.21 (d, 2H),7.19-7.15 (m, 1H), 7.04-6.98 (m, 3H), 5.04 (sept, 1H), 4.88-4.82 (m,1H), 4.58 (d, 1H), 4.40 (d, 1H), 3.89 (s, 1H), 3.87 (s, 3H), 3.17-3.03(m, 2H), 3.09 (s, 3H), 3.00 (s, 3H), 1.25 (d, 3H), 1.23 (d, 3H), 1.15(s, 3H), 1.08 (s, 3H).

¹³C NMR (CDCl₃): δ 170.3, 168.3, 160.2, 154.9, 150.7, 136.9, 132.9,130.5, 130.4, 121.7, 120.2, 120.0, 112.6, 73.4, 69.6, 55.7, 54.5, 53.2,50.4, 37.7, 36.6, 36.3, 29.1, 23.7, 21.6, 21.5.

Example 211 Synthesis ofN-(2-Methoxybenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 by substitution of the appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.92 (dd, 1H), 7.54 (dd, 1H), 7.21 (d, 2H), 7.07-7.00(m, 4H), 6.96 (d, 1H), 5.01 (sept, 1H), 4.83-4.76 (m, 1H), 4.73 (d, 1H),4.61 (d, 1H), 4.17 (s, 1H), 3.93 (s, 3H), 3.14-3.03 (m, 2H), 3.09 (s,3H), 3.00 (s, 3H), 1.36 (s, 3H), 1.22 (d, 3H), 1.21 (s, 3H), 1.19 (d,3H).

¹³C NMR (CDCl₃): δ 170.3, 168.7, 157.7, 154.9, 150.6, 135.4, 33.0,132.5, 30.3, 125.2, 121.7, 120.5, 112.6, 73.3, 69.5, 56.0, 54.8, 53.3,50.4, 37.7, 36.6, 36.3, 29.2, 24.1, 21.6, 21.5.

Example 212 Synthesis of N-(3,4-Dimethoxybenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 by substitution of the appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.50 (dd, 1H), 7.31 (d, 1H), 7.21 (d, 2H), 7.05-7.01(m, 3H), 6.97 (d, 1H), 5.04 (sept, 1H), 4.89-4.82 (m, 1H), 4.56 (d, 1H),4.40 (d, 1H), 3.95 (s, 3H), 3.94 (s, 3H), 3.89 (s, 1H), 3.17-3.03 (m,2H), 3.09 (s, 3H), 3.00 (s, 3H), 1.25 (d, 3H), 1.22 (d, 3H), 1.16 (s,3H), 1.14 (s, 3H).

¹³C NMR (CDCl₃): δ 170.3, 168.5, 154.9, 153.5, 150.7, 149.4, 132.9,130.4, 127.6, 122.3, 121.7, 110.6, 110.3, 73.5, 69.6, 56.3, 56.1, 54.6,53.2, 50.5, 37.7, 36.6, 36.3, 29.2, 23.8, 21.6, 21.5.

Example 213 Synthesis ofN-(2,4-Difluorobenzenesulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 49 by substitution of the appropriatestarting material.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.89 (m, 1H), 7.16 (m, 2H), 6.97 (m, 4H), 6.77 (d,1H), 4.72 (m, 1H), 4.60 (m, 1H), 3.92 (m, 1H), 3.29 (m, 1H), 3.09 (m,5H), 2.93 (s, 3H), 2.70 (m, 2H), 2.55 (m, 1H), 2.10 (m, 1H), 1.42 (s,9H).

¹³C NMR (CDCl₃): δ 170.0, 168.0, 167.7, 137.1, 164.4, 164.3, 161.1,160.9, 157.7, 157.5, 154.8, 150.5, 132.7, 132.6, 132.4, 130.4, 124.0,123.8, 121.7, 112.2, 111.9, 106.5, 106.1, 105.8, 82.6, 55.4, 53.9, 43.5,36.4, 36.2,27.7, 26.8, 25.5.

Example 214 Synthesis ofN-(3,4-Dichlorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 208 usingthe procedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ 8.04 (m, 1H), 7.68 (m, 2H), 7.52 (m, 1H), 7.21 (d,2H), 7.02 (d, 2H), 5.22 (m, 1H), 4.63 (m, 1H), 4.22 (m, 1H), 3.71 (m,1H), 3.57 (m, 1H), 3.30 (m, 3H), 3.08 (s, 3H), 3.02 (m, 3H), 2.97 (s,3H).

¹³CNMR (CD₃OD): δ 174.0, 168.0, 156.9, 152.1, 140.7, 139.3, 135.2,133.2, 131.6, 130.7, 128.3, 123.2, 57.2, 54.9, 54.6, 51.7, 51.4, 43.3,37.3, 36.9, 36.7.

Example 215 Synthesis ofN-(3-Chlorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 207 usingthe procedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD): δ 7.94 (m, 1H), 7.77 (m, 2H), 7.58 (m, 1H), 7.46 (d,1H), 7.19 (d, 2H), 7.07 (d, 2H), 5.23 (m, 1H), 4.63 (m, 1H), 4.20 (m,1H), 3.71 (m, 1H), 3.43 (m, 1H), 3.26 (m, 4H), 3.17 (s, 3H), 2.95 (m,5H).

¹³C NMR(CD₃OD): δ 168.0, 152.1, 142.5, 136.8, 135.0, 132.7, 131.6,128.6, 127.1, 123.3, 57.2, 54.9, 51.4, 51.2, 43.2, 37.2, 36.8, 36.7.

Example 216 Synthesis ofN-(3-Chloro-4-fluorobenzenesulfonyl)-L-(1,1-dioxothiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared using the procedure described in Example92 by substitution of the appropriate starting materials.

NMR data was as follows:

¹H NMR (CD₃OD): δ 7.93 (d), 7.90 (m), 7.29 (s), 7.27 (d), 7.04 (d), 4.60(m), 4.46 (d), 3.90-3.40 (m), 3.10 (s), 2.98 (s), 1.43 (s).

¹³C NMR (CD₃OD): δ 171.5, 166.5, 156.9, 151.9, 135.2, 131.3, 129.9,127.9, 127.8, 123.1, 117.8, 117.5, 101.4, 83.7, 57.9, 56.0, 42.9, 37.3,36.9, 36.7, 28.1.

Example 217 Synthesis ofN-(1-Methylpyrazole4-sulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedures described forthe preparation of Examples 49 and 117.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.77 (s), 7.63 (s), 7.08 (d), 6.93 (d), 6.76 (d), 6.71(d), 5.50 (d), 5.22 (s), 4.82 (t), 4.61 (q), 3.83 (s), 3.25 (dt), 3.04(m), 2.90 (s), 2.05 (dd), 1.34 (s).

¹³C NMR (CDCl₃): δ 169.3, 166.8, 154.7, 150.4, 138.4, 132.4, 132.2,130.2, 121.4, 118.3, 105.4, 82.5, 55.2, 53.6, 53.3, 39.5, 38.3, 36.6,36.3, 36.1, 27.6, 23.5.

Example 218 Synthesis ofN-(3,4-Difluorobenzenesulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 49 by substitution of the appropriatestarting materials.

NMR data was as follows:

¹H NMR (CD₃OD): δ 7.88 (m, 1H), 7.70 (m, 1H), 7.57 (m, 1H), 7.23 (d,2H), 7.03 (d, 2H), 6.83 (d, 1H), 5.63 (dd, 1H), 5.07 (t, 1H), 4.58 (m,1H), 3.22-3.00 (m, 3H), 3.09 (s, 3H), 2.98 (s, 3H), 2.07 (dd, 1H), 1.44(s, 9H).

¹³C NMR (CD₃OD): δ 171.3, 169.3, 156.9, 152.0, 135.0, 131.6, 126.5,122.9, 120.2, 119.9, 119.4, 118.7, 118.4, 106.4, 83.6, 56.5, 55.6, 37.1,36.8, 36.6, 28.1, 25.2.

Example 219 Synthesis ofN-(Toluene-4-sulfonyl)-L-(5,5-dimethyl)thioprolyl-L-(thiomorpholin-4-ylcarbonyloxy)phenylalanineIsopropyl Ester

The title compound was prepared using the procedure described in Example82 by substitution of the appropriate starting materials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 9.09 (s, 1H), 8.88 (m, 1H), 8.16 (m, 1H), 7.50 (m,1H), 7.22 (d, 2H), 7.01 (d, 2H), 6.91 (d, 1H), 5.05 (m, 1H), 4.85 (m,1H), 4.60 (d, 1H), 4.46 (d, 1H), 3.89 (s, 1H), 3.93-3.83 (m, 4H), 3.11(m, 2H), 2.69 (m, 4H), 1.29-1.16 (m, 12H).

¹³C NMR (CDCl₃): δ 170.3, 167.8, 154.3, 153.5, 150.4, 148.7, 135.8,133.1, 132.9, 130.4, 124.0, 121.8, 73.7, 69.7, 54.7, 53.2, 50.5, 47.1,46.4, 37.6, 29.1, 27.4, 27.0, 23.8, 21.6, 21.5.

Example 220 Synthesis ofN-(3,4-Difluorobenzenesulfonyl)-L-(thiamorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 218 usingthe procedure described in Method 11.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.89 (s, 1H), 7.57-7.46 (m, 2H), 7.35 (d, 1H),7.32-7.22 (m, 1H), 7.09 (d, 2H), 6.91 (d, 2H), 6.64 (d, 1H), 5.50 (d,1H), 4.89 (s, 1H), 4.88-4.79 (m, 1H), 3.17-3.02 (m, 3H), 3.02 (s, 3H),2.93 (s, 3H), 1.75 (dd, 1H).

¹³C NMR (CDCl₃): δ 173.6, 167.7, 155.5, 152.0, 151.8, 150.1, 148.4,132.8, 130.4, 124.6, 121.5, 118.7, 118.5, 117.5, 117.3, 117.1, 106.9,54.9, 53.0, 36.4, 36.2, 36.0, 23.4.

Example 221 Synthesis ofN-(2,5-Dichlorothiophene-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title-compound was prepared following the procedure described forthe preparation of Example 2 by substitution of the appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.18 (d, 2H), 7.11 (s, 1H), 7.00 (d, 2H), 6.87 (d,1H), 5.03-4.99 (m, 1H), 4.84-4.81 (m, 1H), 4.65-4.56 (m, 2H), 4.07 (s,1H), 3.10-3.01 (m, 5H), 2.98 (s, 3H), 1.37 (s, 3H), 1.22 (s, 3H), 1.21(s, 3H), 1.18 (s, 3H).

¹³C NMR (CDCl₃): δ 170.3, 167.7, 154.9, 150.7, 132.9, 132.8, 131.9,130.3, 128.0, 127.0, 121.8, 73.4, 69.6, 54.8, 53.2, 50.5, 37.5, 36.6,36.3, 29.1, 23.8, 21.6, 21.5.

Example 222 Synthesis ofN-(1-Methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L4-(thiomorpholin-4-ylcarbonyloxy)phenylalanineIsopropyl Ester

The title compound was prepared using the procedure described in Example82 by substitution of the appropriate starting materials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.89 (s, 1H), 7.80 (s, 1H), 7.21 (d, 2H), 7.01 (m,3H), 5.03 (m, 1H), 4.83 (m, 1H), 4.54 (d, 1H), 4.40 (d, 1H), 3.95 (s,3H), 3.86 (m, 4H), 3.80 (s, 1H), 3.09 (m, 2H), 2.68 (m, 4H), 1.28 (s,3H), 1.22 (m, 6H), 1.16 (s, 3H).

¹³C NMR (CDCl₃): δ 170.4, 168.3, 153.5, 150.4, 139.3, 133.3, 132.9,130.4, 121.7, 117.6, 73.8, 69.7, 54.8, 53.2, 50.5, 47.1, 46.4 39.6,37.6, 29.0, 27.4, 27.1, 24.0, 21.6, 21.5.

Example 223 Synthesis ofN-(8-Quinolinesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 by substitution of the appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 9.01-8.99 (m, 1H), 8.56-8.53 (m, 1H), 8.27-8.23 (m,1H), 8.07-8.04 (m, 1H), 7.66-7.61 (m, 2H), 7.55-7.51 (m, 1H), 7.17 (d,2H), 7.01 (d, 2H), 5.27-5.23 (m, 1H), 5.07-4.98 (m, 1H), 4.84-4.76 (m,1H), 3.34-3.20 (m, 3H), 3.06-2.98 (m, 4H), 2.97 (s, 3H), 2.15-2.09 (m,1H), 1.64-1.51 (m, 3H), 1.23 (d, 6H).

¹³C NMR (CDCl₃): δ 172.0, 170.5, 154.9, 151.5, 150.6, 143.9, 136.8,135.6, 134.9, 134.1, 133.3, 130.2, 129.2, 125.6, 122.3, 121.7, 69.3,62.8, 53.5, 48.7, 37.3, 36.5, 36.3, 29.7, 24.3, 21.6, 21.6.

Example 224 Synthesis ofN-(8-Quinolinesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 223 usingthe procedure described in Method 7.

NMR data was as follows:

¹H NMR (CD₃OD): δ 9.03-9.01 (m, 1H), 8.49-8.42 (m, 2H), 8.23-8.20 (m,1H), 8.09-8.07 (m, 1H), 7.73-7.61 (m, 2H), 7.25 (d, 2H), 7.00 (d, 2H),5.30-5.27 (m, 1H), 4.73-4.69 (m, 1H), 3.38-3.21 (m, 3H), 3.09-3.02 (m,4H), 2.95 (s, 3H), 1.86 (m, 1H), 1.78-1.73 (m, 1H), 1.58-1.50 (m, 2H).

¹³C NMR (CD₃OD): δ 175.3, 174.2, 164.7, 156.9, 152.9,145.2, 138.5,136.9, 135.8, 135.6, 131.6, 130.9, 126.9, 123.8, 122.9, 63.9, 54.7,50.0, 37.5, 36.8, 36.7, 31.6, 25.5.

Example 225 Synthesis ofN-(8-Quinolinesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsoproplyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 by substitution of the appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 9.05-9.03 (m, 1H), 8.53-8.49 (m, 1H), 8.26-8.22 (m,1H), 8.08-8.05 (m, 1H), 7.65-7.60 (m, 1H), 7.56-7.52 (m, 1H), 7.19 (d,2H), 7.06 (d, 1H), 7.00 (d, 2H), 5.17 (d, 1H), 4.94 (m, 1H), 7.74-4.78(m, 2H), 4.66 (s, 1H), 3.08-2.99 (m, 8H), 1.20-1.16 (m, 12H).

¹³C NMR (CDCl₃): δ 170.2, 168.9, 154.9, 151.5, 150.6, 144.2, 136.7,134.4, 134.4, 133.1, 130.3, 129.2, 125.5, 122.3, 121.7, 73.2, 69.3,54.8, 53.3, 50.6, 37.6, 36.6, 36.3, 29.2, 24.1, 21.5, 21.4.

Example 226 Synthesis ofN-(8-Quinolinesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 225 usingthe procedure described in Method 7.

NMR data was as follows:

¹H NMR (CD₃OD): δ 9.06-9.04 (m, 1H), 8.45-8.39 (m, 2H), 8.23-8.14 (m,1H), 7.72-7.61 (m, 2H), 7.32 (d, 2H), 7.03 (d, 2H), 5.12 (d, 1H), 4.87(d, 1H), 4.69-4.64 (m, 2H), 3.28-3.02 (m, 5H), 2.98 (s, 2H), 1.18 (s,3H), 1.08 (s, 3H).

¹³C NMR (CD₃OD): 174.1, 171.8, 157.1, 152.9, 152.0, 145.5, 138.4, 137.3,135.8, 135.6, 135.1, 131.8, 130.9, 126.8, 123.8, 122.9, 73.7, 55.9,54.8, 51.7, 37.6, 36.8, 36.7, 30.2, 25.0.

Example 227 Synthesis ofN-(3—Sulfonamido-4-chloro-benzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 8.45 (d, 1H), 7.91 (d, 1H), 7.67 (d, 1H), 7.13 (d,2H), 7.06 (d, 1H), 7.01 (d, 2H), 5.90 (brs, 2H), 5.06-5.02 (m, 1H),4.79-4.72 (m, 1H), 4.14-4.10 (m, 1H), 3.42-3.39 (m, 1H), 3.25-3.14 (m,2H), 3.07 (s, 3H), 3.04-2.97 (m, 1H), 2.96 (s, 3H), 1.98-1.96 (m, 1H),1.72-1.62 (m, 3H), 1.28-1.25 (in, 6H).

¹³C NMR (CDCl₃): δ 170.8, 170.7, 155.1, 150.6, 141.4, 136.9, 136.1,132.9, 132.8, 131.9, 130.3, 128.7, 121.9, 69.8, 62.1, 53.3, 49.6, 36.9,36.6, 36.4, 30.4, 24.3, 21.6, 21.6.

Example 228 Synthesis ofN-(Toluene-4-sulfonyl)-L-(1-oxothiomorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 182 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.77 (d, 2H), 7.72 (d, 2H), 7.33 (m, 2H), 7.20 (m,2H), 7.12 (d, 2H), 7.01 (m, 2H), 5.10 (m, 1H), 5.01 (m, 1H), 4.84 (m,1H), 4.75 (m, 1H), 3.80 (m, 3H), 3.05 (m, 4H), 2.96 (m, 3H), 2.74 (m,1H), 2.42 (m, 4H), 1.30-1.20 (m, 6H).

¹³C NMR (CDCl₃): δ 170.6, 170.4, 166.8, 166.7, 154.9, 150.7, 150.6,145.1, 144.8, 135.8, 135.5, 132.7, 130.6, 130.4, 130.3, 130.0, 127.7,127.1, 122.4, 121.8, 69.8, 69.4, 55.8, 53.7, 52.9, 50.8, 48.2, 47.9,42.0, 41.2, 38.4, 36.6, 36.5, 36.3, 31.2, 21.5, 21.5.

Example 229 Synthesis ofN-(2,4-Difluorobenzenesulfonyl)-L-(1-oxothiomorpholin-3-carbonyl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 182 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.91 (m, 1H), 7.30 (m, 2H), 6.97 (m, 4H), 4.71 (m,1H), 4.55 (m, 1H), 3.90 (m, 2H), 3.77 (m, 1H), 3.11 (m, 4H), 2.85 (m,3H), 2.80 (m, 1H), 2.60 (m, 2H), 1.46 (s, 9H), 1.39 (s, 9H).

¹³C NMR (CDCl₃): δ 170.0, 168.0, 167.9, 166.4, 166.2, 164.6, 164.4,162.7, 161.4, 161.2, 157.9, 157.8, 154.8, 150.6, 150.4, 132.8, 132.5,132.4, 130.9, 130.4, 130.1, 123.3, 123.1, 122.2, 121.6, 121.1, 122.6,122.2, 111.9, 106.6, 106.3, 105.9, 82.8, 82.3, 55.8, 54.1, 53.2, 51.6,49.2, 48.7, 43.1, 42.3, 38.7, 36.5, 36.2, 31.8, 27.7.

Example 230 Synthesis ofN-(1-Methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine2,2-Dimethylpropyl Ester

The product from Example 161 (1 g., 0.72 mmol) was dissolved inneopentyl alcohol (5 mL). Titanium (IV) isopropoxide (260 mg., 0.9 mmol)was added and the mixture heated at 100° C. under an inert atmospherefor 48 h. Excess neopentyl alcohol was removed under reduced pressureand the residue purified by flash column chromatography (silica, 1% MeOHin CHCl₃) to give the title compound as a white solid (1.02 g, 97%).

Physical data was as follows:

MS (+) ESI [M+H]⁺ 610; [M+NH4]⁺ 627 (100%).

Anal. Calcd. For C₂₉H₃₉N₅O₇S: C, 53.18; H, 6.45; N, 11.49.

Found: C, 53.46; H, 6.38; N, 11.06.

Example 231 Synthesis ofN-(Pyridine-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine2,2-Dimethylpropyl Ester

The product from Example 173 was subjected to the transesterificationprocedure described for the preparation of Example 230. The compound waspurfied by flash column chromatography (silica, 1% MeOH in CHCI₃)followed by recrystallization from ethyl acetate to give the titlecompound as a white solid (720 mg, 47%).

Physical data was as follows:

Anal. Calcd. For C₂₈H₃₈N₄O₇S₂: C, 55.43; H, 6.31; N, 9.23.

Found: C, 55.37; H, 6.32; N, 9.22.

Example 232 Synthesis ofN-(1-Methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineCyclopropylmethyl Ester

The product from Example 161 was subjected to the transesterificationprocedure described for the preparation of Example 230. The titlecompound was obtained as a white solid following flash columnchromatography (silica, 1% MeOH in CHCI₃) (860 mg, 70%).

Physical data was as follows:

Anal. Calcd. For C₂₆H₃₅N₅O₇S₂: C, 52.6; H, 5.94; N, 11.8.

Found: C, 52.49; H, 5.93; N, 11.62.

Example 233 Synthesis ofN-(1-Methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineMethyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 161 and substitution of appropriate startingmaterials.

Physical data was as follows:

MS (+) ESI [M+H]⁺ 554; [M+NH₄]⁺ 571 (100%).

Anal. Calcd. For C₂₃H₃]N₅O₇S₂ 0.2 EtOAc: C, 50.04; H, 5.75; N, 12.26.

Found: C, 50.12; H, 5.69; N, 12.19.

Example 234 Synthesis ofN-(Pyridine-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineEthyl Ester

The product from Example 173 was subjected to the transesterificationprocedure described for the preparation of Example 230. The compound waspurified by flash column chromatography (silica, 2% MeOH in CHCl₃),followed by recrystallization from ethyl acetate to give the titlecompound as a white solid (1.2 g, 61%).

Physical data was as follows:

Anal. Calcd. For C₂₅H₃₂N₄O₇S₂: C, 53.18; H, 5.71; N, 9.92.

Found: C, 53.14; H, 5.72; N, 9.57.

Example 235 Synthesis ofN-(Pyridine-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineCyclopropylmethyl Ester

The product from Example 173 was subjected to the transesterificationprocedure described for the preparation of Example 230. The compound wasisolated as a white solid following flash column chromatography (silica,2% MeOH in CHCl₃) and recrystallization from EtOAc/hexanes (1 g, 65%).

Physical data was as follows:

Anal. Calcd. For C₂₇H₃₄N₄O₄S₂: C, 54.9; H, 5.8; N, 9.48.

Found: C, 54.77; H, 5.65; N, 9.46.

Example 236 Synthesis ofN-(1-Methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L4-(N,N-dimethylcarbamyloxy)phenylalanine2-Methoxyphenyl Ester

To a solution of the compound from Example 139 (1.79 g, 3.31 mmol),2-methoxy-phenol (0.45 g, 3.64 mmol) and BOP (1.61 g, 3.64 mmol) inmethylene chloride (25 mL) at 0° C. was added triethylamine (0.7 mL,4.97 mmol). The reaction mixture was then slowly warmed to 25° C. whereit was stirred, under nitrogen, for 24 h. The reaction was quenched byaddition of 100 mL saturated brine and extracted with EtOAc. The organicextract was washed sequentially with 2N HCl (3 times), saturated sodiumbicarbonate (3×) and saturated brine (2×), dried over MgSO₄, andevaporated to 2.1 g of crude product. Flash chromatography (eluant: 96-4methylene chloride:EtOAc) afforded 1.85 g of a white solid which upontrituration with hexane gave 1.68 g (79%) of white crystals, mp 72-75°C.

Physical data was as follows:

Anal. Calcd. For C₂₉H₃₅N₅O₈S₂: C, 59.94; H, 5.46; N, 10.85.

Found: C, 53.45; H, 5.62; N, 10.31.

Example 237 Synthesis ofN-(1-Methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninen-Butyl Ester

A solution of the compound of Example 139 (2 g) in n-butanol (50 mL) wassaturated upon ice-cooling with HCl gas. The mixture was stirred atambient temperature for 36 h, evaporated in vacuo to almost dryness,then partitioned between 5% NaHCO₃ and chloroform. The organic layer wasdried and evaporated in vacuo to furnish 900 mg of the title compound.

Physical data was as follows:

MS: [(+)ESI], [M+H]⁺ 596.

Example 238 Synthesis ofN-(1-Methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninen-Propyl Ester

A solution ofN-(1-methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine(2 g) in n-propanol (50 mL) was saturated upon ice-cooling with HCl gas.The mixture was stirred at ambient temperature for 36 hours, evaporatedin vacuo to almost dryness, then partioned between 5% NaHCO₃ andchloroform. The organic layer was dried and evaporated in vacuo toprovide 1500 mg of the title compound.

Physical data was as follows:

MS: [(+)ESI], [M+H]⁺ 582.

Example 239 Synthesis ofN-(1-Methylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine2,2-Dimethylpropionyloxymethyl Ester

Potassium iodide (324 mg) was added at once to a mixture of the compoundof Example 139 (1.08 g), chloromethylpivalate (294 mg) and powderedK₂CO₃ (222 mg) in DMF (5 mL). The reaction mixture was stirred atambient temperature overnight, partitioned between water (12 mL) andethyl acetate (60 mL). The separated organic layer was washed with icecold 0.1 N sodium thiosulfate, water and brine, then dried over MgSO₄,filtered and evaporated in vacuo to yield 750 mg of the title compound.

Physical data was as follows:

MS: [(+)ESI], [M+H]⁺ 654.

Example 240 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(4-phenylpiperazin-1-ylcarbonyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure outlined forExample 4 by substitution of the appropriate starting materials. A whitesolid was obtained, mp. 60-65° C.

Physical data was as follows:

MS (+ESI) 694.3 [M+NH4]⁺.

Anal. Calcd. for C₃₆H44N₄O₇S.0.5C₄H₈O₂: C, 63,31, H, 6.71, N, 7.77.

Found: C, 63.12; H, 6.58; N, 7.69.

Example 241 Synthesis ofN-(Toluene4-sulfonyl)-L-prolyl-L-4-(4′-(ethoxycarbonyl)piperidin-1-ylcarbonyloxy)phenylalaninetert-Butyl Ester

The carbamate was prepared by treatment of Tos-Pro-Tyr-t-butyl esterwith 4- nitrophenylchloroformate, followed by addition ofethylisonipecotate (triethylamine, methylene chloride, chilled to 0° C.,then stirred at room temperature overnight). The crude product waspurified by flash chromatography (silica, 95:5 EtoAc:Et₃N) to afford awhite solid. (0.78 g, 39%).

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHz): δ 8.15 (d, 1H, J=7.68 Hz); 7.70 (d, 2H,J=8.34 Hz); 7.40 (d, 2H, J=7.90 Hz); 7.22 (d, 2H, J=8.56 Hz); 7.00 (d,2H, J=8.56 Hz); 4.37 (m, 1H), 4.07 (q, 2H, J=7.14, 14.08 Hz); 4.03 (m,2H); 3.90 (m, 1H); 3.34 (m, 1H); 3.09 (m, 2H); 3.00 (m, 3H); 2.59 (m,1H); 2.39 (s, 3H); 1.87 (m, 2H); 1.58 (m, 5H); 1.41 (m, 1H); 1.35 (s,9H); 1.18 (t, 3H, 7.14 Hz).

IR (KBr, cm⁻¹): 3410, 2990, 2950, 1725, 1680, 1510, 1430, 1355, 1220,1200, 1170, 1000, 675, 595.

MS ((+)ESI, m/z (%)) 689 (100[M+NH₄]⁺); 691 (37[M+NH₄]⁺).

Anal. Calcd. for C₃₄H₄₅N₃O₉S: C, 60.79; H, 6.75; N, 6.25.

Found: C, 60.59; H, 6.67; N, 6.22.

Example 242 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(N-(4′-(2′-aminoethyl)morpholino)carbamyloxy)phenylalanine

The title compound was prepared from the product of Example 152 usingthe procedure described in Method 11.

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHz): δ 12.75 (s, 1H); 8.08 (d, 1H); 7.68 (d, 2H);7.60 (t, 1H); 7.39 (d, 2H); 7.21 (d, 2H); 6.97 (d, 2H); 4.46 (m, iH);4.08 (m, 1H); 3.56 (m, 4H); 3.26 (m, 3H); 3.09 (m, 2H); 2.94 (m, 1H);2.49 (s, 6H); 2.48 (s, 3H); 1.5 (m, 3H); 1.38 (m, 1H).

IR (KBr, cm⁻¹) 3400, 2975, 1725, 1650, 1500, 1350, 1150, 650, 575, 550.

MS ((−)ESI, m/z (%)) 587 (100[M−H]⁺).

Anal. Calcd. for C₂₈H₃₆N₄O₈S.HCOOH.0.5H₂O: C, 54.11; H, 6.11; N, 8.70.

Found: C, 53.96; H, 6.02; N, 8.68.

Example 243 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-[4-(carboxy)piperidin-1-ylcarbonyloxylphenylalanine

The title compound was prepared from the product of Example 241 usingthe procedures described in Methods 6 and 11.

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHz): δ 12.50 (bs, 2H); 8.08 (d, 1H, J=7.90Hz);7.69 (d, 2H, J=8.34 Hz); 7.39 (d, 2H, J=7.90 Hz); 7.22 (D, 2H, J=8.56Hz); 6.99 (d, 2H, J=8.56 Hz); 4.46 (m, 1H); 4.09 (m, 1H); 4.00 (m, 1H);3.90 (m, 1H); 3.30 (m, 1H); 3.09 (m, 3H); 2.95 (m, 2H); 2.49 (m, 1H);2.38 (s, 3H); 1.86 (m, 2H); 1.36-1.61 (m, 6H).

IR (KBr,cm⁻¹) 3400, 2960, 1720, 1535, 1430, 1350, 1200, 1160, 670, 590,550.

MS ((+)ESI, m/z (%)) 605 (100[M+NH₄]⁺).

Anal. Calcd. for C₂₈H₃₃N₃O₉S H₂O: C, 55.53; H, 5.65; N, 6.94.

Found: C, 55.23; H, 5.82; N, 6.59.

Example 244 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(N,N-bis-(2-hydroxyethyl)carbamyloxy)phenylalanineIsopropyl Ester

The carbamate was prepared by treatment of Tos-Pro-Tyr-iPr ester with4-nitrophenyl chloroformate, followed. by addition of diethanol amine(triethylamine, methylene chloride, chilled to 0° C., stirred at roomtemperature overnight). The crude product was purified by flashchromatography (silica, 98:2 EtOAc:EtOH) to afford a white foam.(0.180g, 28%).

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHz): δ 8.26 (d, 1H, J=7.90 Hz); 7.69 (d, 2H,J=8.12 Hz); 7.40 (d, 2H, J=8.12 Hz); 7.23 (D, 2H, J=8.56 Hz); 6.99 (d,2H, J=8.56 Hz); 4.87 (m, 1H); 4.83 (t, 1H, J=5.49 Hz); 4.76 (t, 1H,J=5.49 Hz); 4.42 (m, 1H); 4.08 (m, 1H); 3.58 (m, 2H); 3.51 (m, 2H); 3.44(m, 2H); 3.34 (m, 3H); 2.99-3.09 (m, 3H); 2.39 (s, 3H); 1.59 (m, 3H);1.41 (m, 1H); 1.16 (d, 3H, J=6.15 Hz); 1.12 (d, 3H, J=6.15 Hz).

IR (KBr, cm⁻) 3420, 2940, 1725, 1535, 1670, 1520, 1460, 1410, 1350,1220, 1160, 1110, 670, 600, 550.

MS ((+)ESI, m/z (%)) 606 (15[M+H]⁺); 623 (100[M+NH₂]⁺).

Anal. Calcd. for C₂₉H₃₉N₃O₉S H₂O: C, 56.66; H, 6.56; N, 6.84.

Found: C, 56.66; H, 6.41; N, 6.72.

Example 245 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-[3-(hydroxymethyl)piperidin-1-ylcarbonyloxy]phenylalanineIsopropyl Ester

The carbamate was prepared by treatment of Tos-Pro-Tyr-iPr ester with4-nitrophenyl chloroformate, followed by addition of 3-piperidinemethanol (triethylamine, methylene chloride, chilled to 0° C., stirredat room temperature overnight). The crude product was purified by flashchromatography (silica, 3:2 EtOAc:Hex) to afford a white foam (0.519 g,67%).

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHz): δ 8.26 (d, 1H, J=7.90 Hz); 7.69 (d, 2H,J=8.12 Hz); 7.40 (d, 2H, J=8.12 Hz); 7.22 (d, 2H, J=8.56 Hz); 6.98 (d,2H, J=8.34 Hz); 4.85 (M, 1H); 4.57 (bs, 1H); 4.42 (m, 1H); 3.99-4.09 (m,3H); 3.85 (m, 1H); 3.31 (m, 1H); 3.22 (m, 1H); 2.91-3.10 (m, 4H); 2.80(m, 1H); 2.55 (m, 1H); 2.39 (s, 3H); 1.51-1.72 (m, 6H); 1.42 (m, 2H);1.16 (d, 3H, J=6.15 Hz); 1.11 (d, 3H), J=6.15 Hz).

IR(KBr, cm⁻¹) 3400, 2990, 2940, 2880, 1725, 1520, 1430, 1350, 1220,1165, 1100, 660, 600, 550.

MS ((−))ESI, m/z (%)) 614 (30[M−H]).

Anal. Calcd. for C₃₁H₄₁N₃O₈S: C, 60.47; H, 6.71; N, 6.82.

Found: C, 59.83; H, 6.61; N, 6.59.

Example 246 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(4-trifluoromethanesulfonylpiperazin-1-ylcarbonyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure outlined forExample 128 and substitution of appropriate starting materials.

Physical data was as follows:

MS (+ESI):733 [M+H]⁺.

Anal. Calcd. for C₃lH₃₉F₃N₄O₉S₂.0.10 C₄H₈O₂: C,50.20; H, 5.40; N, 7.55.

Found: C, 50.25; H, 5.46; N, 7.07.

Example 247 Synthesis ofN-(4-(N-Phenylurea)benzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

A mixture of Example 107 (250 mg, 0.51 mmol), phenyl isocyanate (62 mg,0.56 mmol) and triethylamine (76 μl, 0.56 mmol) was heated to refluxunder argon. Reflux was continued overnight. Solvent was removed underreduced pressure and the residue purified by flash chromatography.(silica, hexanes: EtOAc 1:1 then EtOAc) to give the title compound as anoff-white foam (160 mg, 46%), mp 112-115° C.

Physical data was as follows:

MS (+ESI) [M+NH₄]⁺ 697 (100%).

Example 248 Synthesis ofN-(2-Trifluoroacetyl-1,2,3,4-tetrahydroisoquinolin-7-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.70-7.66 (m, 2H), 7.35-7.30 (m, 1H), 7.27-7.21 (m,1H), 7.14-7.10 (m, 2H), 7.01 (d, 2H), 5.09-4.95 (m, 1H), 4.89-4.75 (m,2H), 4.14-4.07 (m, 1H), 3.93-3.85 (m, 2H), 3.35-3.20 (m, 2H), 3.13-2.97(m, 9H), 2.05-2.01 (m, 1H), 1.63 (1.50 (m, 3H), 1.20 (d, 6H).

¹³C NMR (CDCl₃): δ 170.7, 170.6, 170.5, 156.3, 155.8, 154.9, 150.6,140.1, 139.2, 135.1, 135.1, 13.2, 133.0, 133.0, 132.9, 130.2, 130.1,129.9, 126.9, 126.4, 126.3, 125.8, 121.7, 118.3, 114.5, 69.6, 62.1,62.0, 53.2, 49.6, 46.6, 46.5, 45.1, 42.7, 40.9, 37.1, 36.6, 36.3, 30.1,30.0, 29.2, 27.8, 24.2, 24.2, 21.6, 21.6.

Example 249 Synthesis ofN-(1-Methylpyrazole-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

Substituting N-methylpyrazole-3-sulfonyl chloride (See European PatentApplication, 095925) and following the method for the preparation ofExample 56, gave the title compound.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.45 (d, 1H), 7.21 (d, 2H), 7.09 (d, 1H), 7.01 (d,2H), 6.71 (d, 1H), 5.03-4.98 (m, 1H), 4.87-4.84 (m, 1H), 4.60-4.59 (m,2H), 4.05 (s, 1H), 3.97 (s, 3H), 3.12-3.01 (m, 5H), 2.98 (s, 3H),1.22-1.15 (m, 12H).

¹³C NMR (CDCl₃): δ 170.3 , 168.3, 154.9, 150.7, 146.7, 133.0,131.9,130.3, 121.7, 108.9, 73.5, 69.5, 54.7, 53.3, 50.7, 39.9, 37.7,36.6, 36.3, 28.8, 24.1, 21.5, 21.5.

Example 250 Synthesis ofN-(1-Methylpyrazole-3-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 249 usingthe procedure described in Method 7.

NMR data was as follows:

¹H NMR (CD₃OD): δ 8.25 (d, 1H), 7.76 (d, 1H), 7.32 (d, 2H), 7.01 (d,2H), 6.70.(d, 1H), 4.74-4.71 (m, 1H), 4.68 (d, 1H), 4.56 (d, 1H), 4.12(s, 1H), 3.97 (s, 3H), 3.24-3.07 (m, 5H), 2.97 (s, 3H), 1.14 (s, 3H),1.13 (s, 3H).

¹³C NMR (CD₃OD): δ 174.1, 171.4, 157.0, 151.9, 148.2, 135.7, 134.2,131.8, 122.9, 109.6, 74.4, 55.6, 55.0, 51.5, 40.0, 37.6, 36.8, 36.7,29.6, 24.8.

Example 251 Synthesis ofN-(Pyridine-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure outlined for thepreparation of Example 56, where 4-pyridinesulfonyl chloride N-oxide wasused in place of 3-pyridinesulfonyl chloride (see Marsais and coworkers,J. Org. Chem. 1987, 52, 1133-1136). Deoxygenation of the N-oxide wasaccomplished using the procedure of Aoyagi and coworkers, Syntlhesis1997, 891.

NMR data was as follows:

1H NMR (CDCl₃): δ 8.89-8.87 (m, 2H), 7.72-7.70 (m, 2H), 7.19 (d, 2H),7.01 (d, 2H), 6.79 (d, 1H), 5.05-5.01 (m, 1H), 4.85-4.82 (m, 1H), 4.58(d, 1H), 4.45 (d, lH), 3.91 (s, 1H), 3.11-3.02 (m, 5H), 2.99 (s, 3H),1.28-1.16 (m, 12H).

¹³C NMR (CDCl₃): δ 170.3, 167.7, 154.9, 151.5, 150.7, 144.2, 132.7,130.3, 121.8, 120.9, 73.6, 69.7, 54.6, 53.1, 50.4, 37.5, 36.6, 36.3,29.1, 23.6, 21.6, 21.5.

Example 252 Synthesis ofN-(Pyridine-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 251 usingthe procedure described in Method 7.

NMR data was as follows:

¹H NMR (CD₃OD): δ 8.78 (d, 2H), 7.42 (d, 1H), 7.69 (d, 2H), 7.35(d,-2H), 7.06 (d, 2H), 4.69-4.61 (m, 3H), 4.16 (s, 1H), 3.25-3.19 (m,1H), 3.13-3.05 (m, 4H), 2.97 (s, 3H), 1.25 (s, 6H).

¹H C NMR (CD₃OD): δ 174.1, 170.5, 157.4, 152.2, 152.0,147.2, 135.8,131.8, 123.1 122.7, 73.9, 55.6, 54.9, 54.4, 37.5, 36.8, 36.7, 30.1,24.8.

Example 253 Synthesis ofN-(Toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N-methyl-N-(2-dimethylaminoethyl)carbamyloxy)phenylalaninetert-Butyl Ester

A solution of the starting acid (500 mg),(2S)-2-amino-3-{4-[(2-dimethylaminoethyl)-methylcarbamoyloxy]phenyl}propionicacid tert-butyl ester (730 mg), HOBt (235 mg), and 4-methylmorpholine(0.87 mL) in DMF (10 mL) was stirred in ice bath at 0° C. 1-[3-(Dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (360 mg) wasadded to the solution. The ice bath was removed after 10 minutes. Thereaction was stirred at room temperature for 3 hours. Ethyl acetate (20mg) was added. The solution was washed with saturated NaHCO₃ solution(30 mL) 2 times, then washed with brine. The solution was dried withMgSO4. The solvent was evaporated in vacuo, and the residue flashchromatographed on silica gel to give 385 mg of the title compound.

Physical data was as follows:

MS: [(+)ESI], [M+H]⁺663.

Example 254 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(N-methyl-N-(2-dimethylaminoethyl)carbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 253 and substitution of appropriate startingmaterials.

Physical data was as follows:

MS: [(+)ESI], [M+H]⁺617.

Example 255 Synthesis ofN-(Toluene-4-sulfonyl)-L-(5,5-dimethyl)thiapropyl-L-4-(N-methyl-N-(2-dimethylaminoethyl)carbamyloxy)phenylalanine

The title compound was prepared from the product of Example 253 usingthe procedure described in Method 11.

Physical data was as follows:

MS: [(+)ESI], [M+H]^(+607.)

Example 256 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(N-methyl-N-(2-dimethylaminoethyl)carbamyloxy)phenylalanine

The title compound was prepared from the product of Example 254 usingthe procedure described in Method 11.

Physical data was as follows:

MS: [(+)ESI], [M+H]⁺561.

Example 257 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 4 and substitution of appropriate startingmaterials, mp: 64-67° C.

Physical data was as follows:

MS: [M+H]⁺699.

Anal. Calcd. for C₃H₄₀C₁FN₄O₇S₂ H₂O: C, 51.90; H, 5.9; N, 7.8.

Found: C, 51.53; H, 5.50; N, 7.62.

Example 259 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(N,N-dimethycarbamyloxy)phenylalanine

The title compound was prepared for the product of Example 258 using theprocedure described in Method 11.

Physical data was as follows:

MS: [M+1]603.

Anal. Calcd. for C₂₄H₂₇FN₃O₇S₂: C, 49.02; H, 4.63; N, 7.15.

Found: C, 49.25; H, 4.89; N, 6.73.

Example 260 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(thiomorphotin-4-ylcarbonyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 82 and substitution of appropriate startingmaterials, mp. 111-114° C.

Physical data was as follows:

MS: +ESI [M+NH4]⁺ 719.

Anal. Calcd. for C₃₀H₃₇ClFN₃O₇S: C, 50.02; H, 5.46; N, 5.8.

Found: C, 50.23; H, 5.10; N, 5.50.

Example 261 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(thiomorpholin4-ylcarbonyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 82 and substitution of appropriate startingmaterials, mp. 77-81 ° C.

Physical data was as follows:

MS: [M+NH₄]⁺ 705.

Anal. Calcd. for C₂₉H₃₅ClFN₃O₇S₃: C, 50.61; H, 5.13; N, 6.1.

Found: C, 50.33; H, 5.07; N, 5.94.

Example 262 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(N,N-dimethylcarbamyloxy)]phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example-2 and substitution of appropriate startingmaterials, mp. 65-69° C.

Physical data was as follows:

MS: [M+NH₄]⁺ 647.

Anal. Calcd. for C₂₇H₃₃C₁₁FN₃O₇S₂: C, 51.46; H, 5.28; N, 6.4.

Found: C, 51.29; H, 5.19; N, 6.50.

Example 263 Synthesis ofN-(Toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 and substitution of appropriate startingmaterials, mp. 68-72° C.

Physical data was as follows:

MS: [M+H]⁺ 626.

Anal. Calcd. for C₂₈H₃₆ClN₃O₇S₂: C, 53.77; H, 5.80; N, 6.71.

Found: C, 53.26; H, 5.8; N, 6.63.

Example 264 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-methylpiperazin-1-ylcarbonyloxy)lphenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 4 and substitution of appropriate startingmaterials.

Physical data was as follows:

MS: [M+H]⁺ 685.

Anal. Calcd. for C₃₀H₃₈ClN₄O₇: C, 52.59; H, 5.59; N, 8.18.

Found: C, 52.09; H, 5.48; N, 7.77.

Example 265 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-3-chloro-4-(N,N-dimethylcarbamyloxy)lphenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe - - - preparation of Example 2 and substitution of appropriatestarting materials.

Physical data was as follows:

MS: [M+H]⁺580.

Anal. Calcd. for C₂₇H₃₄ClN₃O₇S 0.5 H₂0: C, 55.04; H, 6.00; N, 7.13.

Found: C, 55.06; H, 5.71; N, 6.93.

Example 266 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-(2′-pyridyl)-piperazin-1-ylcarbonyloxy)lphenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 4 and substitution of appropriate startingmaterials.

Physical data was as follows:

MS: [M+H]⁺ 748.

Anal. Calcd. for C₃₄H₃₉ClFN₅O₇S₂: C, 54.57; H, 5.25; N, 9.3.

Found: C, 54.26; H, 5.10; N, 9.07.

Example 267 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-3-chloro-4-(4-(2′-pyridyl)-piperazin-1-ylcarbonyloxy)lphenylalanine tert-Butyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 4 and substitution of appropriate startingmaterials, mp. 80-86° C.

Physical data was as follows:

MS: [M+H]⁺ 762.

Anal. Calcd. for C₃₅H₄₁ClFN₅O₇S₂: C, 55.14; H, 5.42; N, 9.19.

Found: C, 54.67; H, 5.40; N, 8.69.

Example 268 Synthesis ofN-(4-Nitrobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared following the procedure described forthe preparation of Example 2 and substitution of appropriate startingmaterials.

Physical data was as follows:

Anal. Calcd. for C₂₆H₃₂N₄O₉S: C, 54.16; H, 5.59; N, 9.72.

Found: C, 53.69; H, 5.24; N, 9.52.

Example 269 Synthesis ofN-(4-Aminobenzenesulfonyl)-L-prolyl-L4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared from the product of Example 268 usingthe procedure described in Method 4.

Physical data was as follows:

Anal. Calcd. for C₂₆H₃₄N₄O₇S: C, 57.13; H, 6.27; N, 10.25.

Found: C, 56.30; H, 6.12; N, 10.05.

Example 270 Synthesis of 5N-(Toluene-4-sulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 82 and substitution of appropriate startingmaterials.

Physical data was as follows:

Anal. Calcd. for C₂₉H₃₇N₃O₇S₂: C, 57.69; H, 6.18; N, 6.96.

Found: C, 57.36; H, 5.99; N, 6.76.

Example 271 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(4-phenylcarbamylpiperazin-1-ylcarbonyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 4 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHz): δ 8.62 (s, 1H); 8.11 (d, 1H); 7.73 (d, 2H);7.45 (m, 4H); 7.26 (m, 3H); 7.04 (m, 2H); 6.95 (m, lH); 6.25 (d, 1H);4.90 (m, 1H); 4.50 (m, 1H); 4.11 (m, 1H); 3.6 (br, 4H); 3.4 (br, 4H);3.10 (m, 2H); 3.00 (m, 1H); 2.40 (s, 3H); 1.60 (m, 3H); 1.40 (m, 1H);1.18 (d, 3H); 1.12 (d, 3H).

IR (KBr, cm⁻¹) 3400-3500(br), 2950, 2900, 1725, 1650, 1540, 1450, 1240,1210, 1000, 760, 675, 580, 540.

MS ((+)ESI, m/z (%)) 706 (100 [M+H]⁺).

Anal. Calcd. for C₃₆H₄₃N₅O₈S.0.35 EtOAc: C, 60.98; H, 6.27; N, 9.51.

Found: C, 50.31; H, 6.16; N, 9.33.

Example 272 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-(4-phenylcarbamylpiperazin-1-ylcarbonyloxy)phenylalanine

The title compound was prepared from the product of Example 271 usingthe procedure described in Method 7.

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHz): δ 12.8 (s, 1H); 8.62 (s, 1H); 8.11 (d, 1H);7.73 (d, 2H), 7.45 (m, 4H); 7.26 (m, 3H); 7.04 (m, 2H); 6.95 (m, 1H);6.25 (d, 1H); 4.50 (m, 1H); 4.11 (m, 1H); 3.6 (br, 4H); 3.4 (br, 4H);3.10 (m, 2H); 3.00 (m, 1H); 2.40 (s, 3H); 1.60 (m, 3H); 1.40 (m, 1H).

IR (KBr, cm⁻¹) 3400, 1725, 1650, 1540, 1450, 1240, 1210, 1000, 760, 675,580, 540.

MS ((−)ESI, nl/z (%)) 662 (100 [M−H]+).

Example 273 Synthesis ofN-(1-n-Butylpyrazole-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 137 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (CDCl₃): δ 7.89 (s, 1H), 7.83 (s, 1H), 7.21 (d, 2H), 7.06 (d,1H), 7.02 (d, 2H), 5.04 (sept, 1H), 4.89-4.82 (m, 1H), 4.57 (d, 1H),4.41 (d, 1H), 4.16 (t, 2H), 3.78 (s, 1H), 3.14 (dd, 1H), 3.06 (dd, 1H),3.09 (s, 3H), 3.00 (s, 3H), 1.85 (pent, 2H), 1.36-1.23 (m, 2H), 1.27 (s,3H), 1.24 (d, 3H), 1.21 (d, 3H), 1.16 (s, 3H).

¹³C NMR (CDCl₃): δ 170.4, 168.3, 154.9, 150.7, 139.2, 131.8, 130.3,121.8, 117.0, 73.8, 69.6, 54.8, 53.2, 52.7, 50.6, 37.7, 36.6, 36.3,31.8, 28.9, 24.0, 21.6, 21.5, 19.4, 13.3.

Example 274 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L4-(pyridin4-ylcarbonyl)piperazin-1-ylcarbonyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 4 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHz): δ 8.69 (dd, 2H), 8.28 (d, 1H); 7.71 (d, 2H);7.43 (m, 4H); 7.26 (d, 2H); 7.04 (d, 2H); 4.86 (m, 1H); 4.42 (m, 1H);4.05 (m, 1H); 3.4-3.8 (brm, 9H); 3.05 (m, 3H); 2.40 (s, 3H); 1.60 (m,3H); 1.40 (m, 1H); 1.18 (d, 3H); 1.15 (d, 3H).

IR (KBr, cm⁻¹) 3400, 1725, 1650, 1510, 1200, 1160, 1100, 1010, 650, 600,550.

MS ((+)ESI, m/z (%)) 692 (100 [M+H]⁺).

Anal. Calcd. for C₃₅H₄₁N₅O₉S. 0.75H₂0: C, 59.60; H, 6.07; N, 9.93

Found: C, 59.45; H, 5.86; N, 9.88.

Example 275 Synthesis ofN-(Toluene-4-sulfonyl)-L-4-oxoprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 164 usingthe procedure described in Method 11.

Physical data was as follows:

MS [(−)ESI] [M−H]) 516.

Example 276 Synthesis ofN-(Toluene-4-sulfonyl)-L-trans-4-hydroxyprolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 165 usingthe procedure described in Method 11.

Physical data was as follows:

MS [(−)ESI] [M−H]) 518.

Example 277 Synthesis ofN-(4-Cyanobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 and substitution of appropriate startingmaterials, mp. 166-167° C.

Example 278 Synthesis ofN-(4-Aminobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 107 usingthe procedure described in Method 11.

Physical data was as follows:

Anal. Calcd. For C₂₃H₂₈N₄O₇S: C, 47.34; H, 4.84;N, 9.60.

Found: C, 47.57; H, 5.20; N, 8.75.

Example 279 Synthesis ofN-(Toluene-4-sulfonyl)-L-4-oxoprolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalaninetert-Butyl Ester

Acetonitrile (3 mL) was cooled to −40° C. (CH₃CN/dry ice). Oxalylchloride (0.10 mL) was added.N-(Toluene-4-sulfonyl)-L-4-hydroxyprolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalaninetert-butyl ester (300 mg) and dry DMSO (0.008 mL) were dissolved inacetonitrile (4 mL) and were added to the above solution. The reactionwas stirred at −40° C. for half an hour under dry conditions.Triethylamine (0.33 mL) was added to the solution. The dry ice bath wasremoved after 5 minutes. The reaction was stirred at room temperaturefor 1 hour. The solvent was evaporated in vacuo, and ethyl acetate (15mL) was added. The mixture was washed with water (3×), then washed withbrine. The solution was dried over MgSO₄. The solvent was evaporated invacuo, and the residue was flushed on a silica gel column to give 150 mgof the title compound, mp. 84-85° C.

Example 280 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L4-[3-(hydroxymethyl)piperidin-1-ylcarbonyloxy]phenylalanine

The title compound was prepared following the procedure described forthe preparation of Example 4 and substitution of appropriate startingmaterials, mp. 84-85° C.

Example 281 Synthesis ofN-(Toluene4-sulfonyl)-L-(4,4-difluoro)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 2 and substitution of appropriate startingmaterials.

Physical data was as follows:

MS: [(+)ESI], [M+NH₄]⁺ 599.

Example 282 Synthesis ofN-(Toluene-4-sulfonyl)-L-(4,4-difluoro)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 281 usingthe procedure described in Method 7.

Physical data was as follows:

MS: [(+)ESI], [M+NH₄]557.

Example 283 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-(4-benzoylpiperazin-1-ylcarbonyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 4 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHz): δ 8.27 (d, 1H); 7.69 (d, 2H); 7.45 (m, 7H);7.24 9d, 2H); 7.02 (d, 2H); 4.86 (m, 1H); 4.42 (m, 1H); 4.07 (m, 1H);3.65 (br s, 4H); 3.45 (br s, 4H); 3.35 (m, 1H); 3.05 (m, 3H); 2.38 (s,3H); 1.60 (m, 3H); 1.40 (m, 1H); 1.18 (d, 3H); 1.11 (d, 3H).

IR(KBr, cm⁻¹) 3400, 1725, 1675, 1625, 1510, 1425, 1350, 1250, 1175,1110, 1010, 700, 660, 590, 550.

MS ((+)ESI, m/z (%)) 708 (100 [M+NH₂]⁺)

Anal. Calcd. for C₃₆H₄₂N₄O₈S.0.5H₂0: C, 61.79; H, 6.19; N, 8.01.

Found: C, 61.64; H, 6.10; N, 7.72.

Example 284 Synthesis ofN-(1-Methyl-1H-imidazole-4-sulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The carbamate was prepared by treatment of 1-methylimidazole-4-sulfonyl-Pro-Try-iPr ester with 4-nitrophenylchloroformate, followed by addition of dimethylalamine (triethylamine,methylene chloride, 0° C., stirred at room temperature overnight.) Thecrude product was purified by flash chromatography (silica, 95:3:2EtOAc:EtOH;Et₃N), followed by recrystallization from EtOAc. A whitesolid was obtained, mp 162-164° C. (8.7g, 66%).

Physical data was as follows:

Anal. Calcd. for C₂₄H₃₃N₅O₇S: C, 53.82; H, 6.21; N, 13.08.

Found: C, 53.47; H, 6.13; N, 12.96.

Example 286 Synthesis ofN-(Toluene-4-sulfonyl)-L-4-(thiomorpholin-4-ylcarbonyloxy)prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine

The title compound was prepared from the product of Example 285 usingthe procedure described in Method 11, mp. 116-118° C.

Example 287 Synthesis ofN-(4-Cyanobenzenesulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 4 and substitution of appropriate startingmaterials, mp. 70-71° C.

Example 288 Synthesis ofN-(4-Amidinobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineMethyl Ester

Methanol (dry) was cooled to 0° C. HCl was bubbled in the solution for15 minutes to make a saturated solution. Example 277 was added and thereaction mixture was stirred at 0° C. for 30 minutes then at roomtemperature for 24 hours. The solvent was evaporated. NH₃ in methanol(2M, 5 mL) was added. The reaction was stirred at room temperature for24 hours. The solvent was evaporated. The residue was purified byreverse phase HPLC in CH₃CN:H₂O (20:80). At a retention time of 12.45minutes, the product was isolated and freeze-dried to provide the titlecompound.

NMR data was as follows:

¹H NMR (in DMSO) multiplet at 1.47-1.55 ppm (1H), 1.63-1.72 ppm (3H′s),singlet at 2.87 ppm (3H′s), singlet at 3.02 ppm (3H′s), multiplet at3.05-3.10 ppm (2H′s), multiplet at 3.17-3.22 ppm (1 H), multiplet at3.37-3.42 ppm (1H), singlet at 3.62 ppm (3H′s), multiplet at 4.21-4.23ppm (1H), quartet at 4.48-4.56 ppm (1H), doublet at 7.00-7.03 ppm(2H′s), doublet at 7.23-7.26 ppm (2H′s), a broad peak at 7.20-7.50 ppm,doublet at 8.02-8.03 ppm (4H′s), doublet at 8.48-8.52 ppm (1H).

Example 289 Synthesis ofN-(Toluene-4-sulfonyl)-L-4-hydroxyprolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 4 and substitution of appropriate startingmaterials, mp. 80-82° C.

Example 290 Synthesis ofN-(Toluene-4-sulfonyl)-L-4-hydroxyprolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine

N-(Toluene-4-sulfonyl)-L-4-hydroxyprolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalaninetert-butyl ester (160 mg) was dissolved in formic acid (7 mL). Thereaction was stirred at room temperature for 6 hours. The solvent wasevaporated and the residue purified using reverse phase HPLC in 20:80CH₃CN/water. At a retention time of 5.85 minutes, 50 mg of the titlecompound was obtained, mp. 170-172° C.

Example 291 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-(4-benzoylpiperazin-1-ylcarbonyloxy)phenylalanine

The title compound was prepared from the product of Example 283 usingthe procedure described in Method 11.

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHz): δ 12.8 (s, 1H); 8.27 (d, 1H); 7.69 (d, 2H);7.45 (m, 7H); 7.24 (d, 2H); 7.02 (d, 2H); 4.42 (m, 1H); 4.07 (m, 1H);3.65 (br s, 4H); 3.45 (br s, 4H); 3.35 (m, 1H); 3.05 (m, 3H); 2.38 (s,3H); 1.60 (m, 3H); 1.40 (m, 1H).

IR (KBr, cm⁻¹) 3400, 1725, 1675, 1625, 1510, 1425, 1350, 1260, 1175,1110, 1010, 700, 660, 590, 550.

MS ((+)ESI, m/z (%)) 666 (100 [M+NH₄]⁺).

Anal. Calcd. for C₃₃H₃₆N₄O₈S.0.66H₂O: C, 60.00; H, 5.69; N, 8.48

Found: C, 60.36; H, 5.70; N, 7.81.

Example 292 Synthesis ofN-(4-Amidinobenzenesulfonyl)-L-prolyl-L4-(thiomorpholin-4-ylcarbonyloxy)phenylalanineMethyl Ester

The title compound was prepared following the procedure described forthe preparation of Examples 287 and 288.

Physical data was as follows:

MS: [(+)ESI] [M+H] 604.

Example 293 Synthesis ofN-(3-Fluorobenzenesulfonyl)-L-prolyl-L4-(N,N-dimethylcarbonyloxy)phenylalanine

The title compound was prepared from the product of Example 166 usingthe procedure described in Method 11, mp. 82-83° C.

Example 294 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-L-4-[N-methyl-N-(2-(1-methyl-toluenesulfonyl-amino]ethyl)carbamyloxylphenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 4 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHz): δ 8.27 (d, 1H); 7.71 (d, 2H); 7.69 (d, 2H);7.40 (m, 4H); 7.24 (d, 2H); 6.99 (d, 2H); 4.86 (m, 1H); 4.43 (m, 1H);4.06 (m, 1H); 3.51 (m, 1H); 3.2-3.35 (m, 3H); 2.9-3.2 (overlapping m,7H); 2.67 (d, 3H); 2.38 (s, 6H); 1.60 (m, 3H); 1.40 (m, 1H); 1.20 (d,3H); 1.15 (d, 3H).

IR (KBr, cm⁻¹) 3400, 2975, 2950, 1725, 1680, 1510, 1450, 1400, 1280,1225, 1150, 1110, 800, 730, 675, 575, 550.

MS ((+)ESI, m/z (%)) 760 (100 [M+NH₄]⁺).

Anal. Calcd. for C₃₆H₄₆N₄O₉S₂: C, 58.20; H, 6.24; N, 7.54.

Found: C, 57.90; H, 6.30; N, 7.34.

Example 295 Synthesis of N-(Toluene-4-sulfonyl)-L-prolyl-L-4-[N-(2-(N′-phenylaminocarbonyloxy)ethyl)carbamyloxy)]phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described forthe preparation of Example 4 and substitution of appropriate startingmaterials.

NMR data was as follows:

¹H NMR (DMSO-d₆, 400 MHz): δ 9.67 (s, 1H); 8.27 (d, 1H); 7.72 (d, 2H);7.47 (d, 2H); 7.42 (d, 2H); 7.24 (m, 4H); 6.98 (m, 3H); 4.87 (m, lh);4.45 (m, 1H); 4.18 (m, 2H); 4.05 (m, 1H); 3.4 (m, 3H); 3.05 (m, 3H) 2.40(s, 3H); 1.6 (m, 3H); 1.40 (m, 1H); 1.2 (d, 3H); 1.15 (d, 3H).

IR (KBr, cm⁻¹) 3350, 2950, 1725, 1675, 1600, 1550, 1500, 1325, 1200,1150, 1100, 650, 575, 525.

MS ((+)ESI, m/z (%)) 698 (100 [M+NH₄]⁺).

Anal. Calcd. for C₃₄H₄₀N₄O₉S.0.21 EtOAc. 0.5H₂O: C, 59.08; H, 6.07; N,7.91.

Found: C, 59.08; H, 6.02; N, 7.80.

Example 296 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-4-(trans-hydroxy)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

The title compound was prepared following the procedure described inExample 2 and substitution of appropriate starting materials.

Physical data was as follows:

MS: [(+)ESI], [M+NH₄]583.

Example 297 Synthesis ofN-(4-Fluorobenzenesulfonyl)-L-4-(trans-hydroxy)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The title compound was prepared following the procedure described inExample 2 and substitution of appropriate starting materials.

Physical data was as follows:

MS: [(+)ESI], [M+NH₄]597.

Example 298 Synthesis ofN-(4-Amidinobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared from the product of Example 288 usingthe procedure described in Method 5, mp. 130-132° C.

Example 299 Synthesis of Piperazine-1,4-dicarboxylic AcidBis-{4-[(2S)-2-tert-butoxycarbonyl-2-((4R)-5,5-dimethyl-3-(toluene-4-sulfonyl)thiazolidine-4-carboxamido)ethyl]phenyl}Ester

The title compound was prepared following the procedure described inExample 4, except that 0.5 equivalents of piperazine were used.

Physical data was as follows:

Anal. Calcd. for C₅₈H₇₄N₆O₁₄S₄: C, 57.69; H, 6.18; N, 6.96.

Found: C, 58.01; H, 6.07; N, 6.68.

Example 300 Synthesis of Piperazine- 1,4-dicarboxylic AcidBis-{4-[(2S)-2-carboxy-2-((4R)-5,5-dimethyl-3-(toluene-4-sulfonyl)thiazolidine-4-carboxamido)ethyl]phenyl}Ester

The title compound was prepared by hydrolysis of the di-t-butyl esterfrom Example 299 with formic acid to give a white foam (300 mg,quantitative).

Physical data was as follows:

Anal. Calcd. for C₅₀H₅₈N₆O₁₄S₄: C, 54.83; H, 5.34; N, 7.67

Found: C, 55.10; H, 5.57; N, 7.37.

Other compounds of Formulae I and Ia prepared by the methods describedabove include those set forth in Examples 301-370 in Table 11 below.TABLE 11

Ex. No. R¹ R² R³ R⁵ R⁶ 301 p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O-]benzyl- —OC(CH₃)₃ —CH₂CH₂—NH—CH₂— (L-piperazinyl) 302p-F-φ- R²/R³ = cyclic p-[(2-(hydroxymethyl)pyrrolidin-1-yl-C(O)O-]-—OC(CH₃)₃ —CH₂—S—C(CH₃)₂— benzyl- (L-5,5- dimethylthiazolidin-4- yl) 303p-F-φ- R²/R³ = cyclic p-[(2-(hydroxymethyl)pyrrolidin-1-yl-C(O)O-]- —OH—CH₂—S—C(CH₃)₂— benzyl- (L-5,5- dimethylthiazolidin-4- yl) 304 p-CH₃-φ-R²/R³ = cyclic p-[(2-(CH₃OC(O)-)pyrrolidin-1-yl)-C(O)O-] —OC(CH₃)₃ 3carbon atoms benzyl (L-pyrrolidinyl) 305 p-F-φ- R²/R³ = cyclic3-chloro-4-[(thiomorpholin-4-yl)-C(O)O-] —OH —CH₂—S—C(CH₃)₂— benzyl-(L-5,5- dimethylthiazolidin-4- yl) 306 p-F-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)-C(O)O-] —OH —CH₂—S—C(CH₃)₂— benzyl-(L-5,5- dimethylthiazolidin-4- yl) 307 p-F-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)-C(O)O-] —OC(CH₃)₃ —CH₂—S—C(CH₃)₂—benzyl- (L-5,5- dimethylthiazolidin-4- yl) 308 p-CH₃-φ- R²/R³ = cyclicp-[(thiomorpholin-4-yl)-C(O)O-]benzyl- —OC(CH₃)₃ —CH₂CH(OH)CH₂— (L-4-hydroxypyrrolidinyl) 309 p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O-]benzyl- —O(CH₂CH₂O)₂CH₃ 3 carbon atoms (L-pyrrolidinyl)310 p-F-φ- R²/R³ = cyclic p-[(4-(pyrimidin-2-yl)piperazin-1-yl)-—OC(CH₃)₃ —CH₂—S—C(CH₃)₂— C(O)O-]benzyl- (L-5,5- dimethylthiazolidin-4-yl) 311 p-F-φ- R²/R³ = cyclic 3-fluoro-4-[(CH₃)₂NC(O)O-]-benzyl-—OCH(CH₃)₂ —CH₂—S—C(CH₃)₂— (L-5,5- dimethylthiazolidin-4- yl) 312p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O-]benzyl- —OC(CH₃)₃ —CH₂—CH₂N—(—SO₂CH₃)—CH₂— (L-4- methanesulfonyl- piperazinyl) 313 R¹/R² = Hp-[(CH₃)₂NC(O)O-]benzyl- —OH 1,1-dioxo-2,3- dihydro-3,3- dimethyl-1,2-benzisothiazol- 2-yl- 314 R¹/R² = H p-[(CH₃)₂NC(O)O-]benzyl- —OC(CH₃)₃N-2,10- camphorsultamyl- 315 R¹/R² = H p-[(CH₃)₂NC(O)O-]benzyl- —OHN-2,10- camphorsultamyl- 316 R¹/R² = H3-chloro-4-[(CH₃)₂NC(O)O-]-benzyl- —OCH(CH₃)₂ N-2,10- camphorsultamyl-317 p-Br-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O-]benzyl- —OC(CH₃)₃—CH₂—S—C(CH₃)₂— (L-5,5- dimethylthiazolidin-4- yl) 318 p-Br-φ- R²/R³ =cyclic p-[(CH₃)₂NC(O)O-]benzyl- —OH —CH₂—S—C(CH₃)₂— (L-5,5-dimethylthiazolidin-4- yl) 319 p-CH₃-φ- R²/R³ = cyclicp-[(4-methylpiperazin-1-yl)-C(O)O-]benzyl- —OH —CH₂—CH(OH)—CH₂— (L-4-hydroxypyrrolidinyl) 320 p-F-φ- R²/R³ = cyclicp-[(4-pyrimidin-2-yl)piperazin-1-yl)-C(O)O-] —OH —CH₂—S—C(CH₃)₂— benzyl(L-5,5- dimethylthiazolidin-4- yl) 321 p-F-φ- R²/R³ = cyclicp-[4-(pyridin-2-yl)piperazin-1-yl)C(O)O-] —OCH(CH₃)₂ —CH₂—S—C(CH₃)₂]benzyl (L-5,5- dimethylthiazolidin-4- yl) 322 p-F-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O-]benzyl- —OH —CH₂—CH₂—S— (thiazolidin-2-yl) 323 p-F-φ-R²/R³ = cyclic p-[(CH₃)₂NC(O)O-]benzyl- —OC(CH₃)₃ —CH₂—CH₂—S—(thiazolidin-2-yl) 324 p-CH₃-φ- R²/R³ = cyclicp-[(thiomorpholin-4-yl)C(O)O-]benzyl- —OH —CH₂—C(O)—CH₂—(L-4-oxopyrrolidinyl) 325 p-CH₃-φ- R²/R³ = cyclicp-[(4-methylpiperazin-1-yl)-C(O)O-]benzyl- —OH —CH₂—C(O)—CH₂—(L-4-oxopyrrolidinyl) 326 p-F-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)- —OH —CH₂—CH₂—S— C(O)O-]benzyl-(thiazolidin-2-yl) 327 p-NO₂-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)- —OC(CH₃)₃ 3 carbon atomsC(O)O-]benzyl- (L-pyrrolidinyl) 328 p-F-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)- —OC(CH₃)₃ —CH₂—CH₂—S—C(O)O-]benzyl- (thiazolidin-2-yl) 329 p-Br-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)- —OH —CH₂—S—C(CH₃)₂— C(O)O-]benzyl-(L-5,5- dimethylthiazolidin-4- yl) 330 p-CH₃-φ- R²/R³ = cyclicp-[(4-(φNHC(S)-)piperazin-1-yl)-C(O)O-] —OCH(CH₃)₂ 3 carbon atomsbenzyl- (L-pyrrolidinyl) 331 p-F-φ- R²/R³ = cyclicp-[(4-CH₃—homopiperazin-1-yl)-C(O)O-]benzyl- —OC(CH₃)₃ —CH₂—CH₂—S—(thiazolidin-2-yl) 332 p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O-]benzyl-—OC(CH₃)₃ —CH₂CH(—OSO₂CH₃)— CH₂— (L-4-methanesulfoxy- pyrrolidinyl) 333p-H₂NC(O)-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O-]benzyl- —OH 3 carbon atoms(L-pyrrolidinyl) 334 p-H₂NC(O)-φ- R²/R³ = cyclicp-[(thiomorpholin-4-yl)C(O)O-]benzyl- —OH 3 carbon atoms(L-pyrrolidinyl) 335 p-H₂NC(═N)-φ- R²/R³ = cyclicp-[(thiomorpholin-4-yl)C(O)O-]benzyl- —OH 3 carbon atoms(L-pyrrolidinyl) 336 p-NO₂-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)-C(O)O-] —OH 3 carbon atoms benzyl(L-pyrrolidinyl) 337 p-F-φ- R²/R³ = cyclic3-chloro-4-[(4-pyridin-2-yl)piperazin-1- —OCH₂CH₃ —CH₂—S—C(CH₃)₂—yl)C(O)O-]- (L-5,5- benzyl- dimethylthiazolidin-4- yl) 338 p-F-φ- R²/R³= cyclic 3-chloro-4-[(4-pyridin-2-yl)piperazin-1- —OH —CH₂—S—C(CH₃)₂—yl)C(O)O-]- (L-5,5- benzyl- dimethylthiazolidin-4- yl) 339 p-F-φ- R²/R³= cyclic p-[(4-CH₃-homopiperazin-1-yl-C(O)O-]benzyl- —OH —CH₂—CH₂—S—(thiazolidin-2-yl) 340 1- R²/R³ = cyclic3-chloro-4-[(CH₃)₂NC(O)O-]benzyl- —OCH(CH₃)₂ methylpyrazol-—CH₂—S—C(CH₃)₂— 4-yl- (L-5,5- dimethylthiazolidin-4- yl) 341 1- R²/R³ =cyclic p-[(4-(pyridin-2-yl)piperazin-1-yl)C(O)O-] —OCH(CH₃)₂methylimidazol- 3 carbon atoms benzyl 4-yl- (L-pyrrolidinyl) 342 1-R²/R³ = cyclic p-[(4-(pyridin-2-yl)piperazin-1-yl)C(O)O-] —OC(CH₃)₃methylimidazol- 3 carbon atoms benzyl 4-yl- (L-pyrrolidinyl) 343p-CH₃-φ- R²/R³ = cyclic p-[(4-(pyridin-2-yl)piperazin-1-yl)C(O)O-] —OH 3carbon atoms benzyl (L-pyrrolidinyl) 344 p-CH₃-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)C(O)O-] —OC(CH₃)₃ 3 carbon atomsbenzyl (L-pyrrolidinyl) 345 p-CH₃-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)-C(O)O-] —OCH(CH₃)₂ 3 carbon atomsbenzyl (L-pyrrolidinyl) 346 p-F-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)C(O)O-] —OCH(CH₃)₂ 3 carbon atomsbenzyl- (L-pyrrolidinyl) 347 p-F-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)C(O)O-] —OC(CH₃)₃ 3 carbon atomsbenzyl- (L-pyrrolidinyl) 348 p-CH₃-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O-]benzyl- —OH —CH₂CH₂N(—SO₂— CH₃)CH₂— (4-methanesulfonyl-piperazin-2-yl) 349 p-CH₃-φ- R²/R³ = cyclic p-[(CH₃)₂NC(O)O-]benzyl- —OH—CH₂CH(—OSO₂— CH₃)CH₂— (L-4-methanesulfoxy- pyrrolidinyl) 350 CH₃— —CH₂φH p-[(CH₃)₂NC(O)O-]benzyl- —OC(CH₃)₃ 351 p-Br-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)C(O)O-] —OC(CH₃)₃ —CH₂—S—C(CH₃)₂—benzyl- (L-5,5- dimethylthiazolidin-4- yl) 352 p-CF₃O-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O-]benzyl- —OH —CH₂—S—C(CH₃)₂— (L-5,5-dimethylthiazolidin-4- yl) 353 p-CF₃O-φ- R²/R³ = cyclicp-[(CH₃)₂NC(O)O-]benzyl- —OC(CH₃)₃ —CH₂—S—C(CH₃)₂— (L-5,5-dimethylthiazolidin-4- yl) 354 p-CF₃O-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)C(O)O-] —OC(CH₃)₃ —CH₂—S—C(CH₃)₂—benzyl- (L-5,5- dimethylthiazolidin-4- yl) 355 p-F-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)C(O)O-] —OH 3 carbon atoms benzyl-(L-pyrrolidinyl) 356 p-F-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)C(O)O-] —OH —CH₂CH(OH)CH₂— benzyl-(L-4- hydroxypyrrolidinyl) 357 p-CF₃O-φ- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)C(O)O-] —OH —CH₂—S—C(CH₃)₂— benzyl-(L-5,5- dimethylthiazolidin-4- yl) 358 1- R²/R³ = cyclic3-chloro-4-[(CH₃)₂NC(O)-]-benzyl- —OH methylimidazol- 3 carbon atoms4-yl- (L-pyrrolidinyl) 359 1- R²/R³ = cyclic3-chloro-4-[(CH₃)₂NC(O)O-]-benzyl- —OCH(CH₃)₂ methylimidazol- 3 carbonatoms 4-yl- (L-pyrrolidinyl) 360 1- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)-C(O)O-] —OH methylimidazol- 3 carbonatoms benzyl- 4-yl- (L-pyrrolidinyl) 361 1- R²/R³ = cyclicp-[(4-(pyridin-2-yl)piperazin-1-yl)C(O)O-] —OH methylimidazol-—CH₂—S—C(CH₃)₂— benzyl- 4-yl- (L-5,5- dimethylthiazolidin-4- yl) 362 1-R²/R³ = cyclic p-[(4-(pyridin-2-yl)piperazin-1-yl)C(O)O-] —OHmethylpyrazol- 3 carbon atoms benzyl 4-yl- (L-pyrrolidinyl) 363 1- R²/R³= cyclic p-[(4-(pyridin-2-yl)piperazin-1-yl)C(O)O-] —OCH(CH₃)₂methylpyrazol- 3 carbon atoms benzyl 4-yl- (L-pyrrolidinyl) 364 1- R²/R³= cyclic p-[(4-(pyridin-2-yl)piperazin-1-yl)C(O)O-] —OC(CH₃)₃methylpyrazol- 3 carbon atoms benzyl 4-yl- (L-pyrrolidinyl) 365 1- R²/R³= cyclic p-[(4-(pyridin-2-yl)piperazin-1-yl)C(O)O-] —OC(CH₃)₃methylpyrazol- —CH₂—S—C(CH₃)₂— benzyl 4-yl- (L-5,5-dimethylthiazolidin-4- yl) 366 1- R²/R³ = cyclic3-chloro-4-[(4-(pyridin-2-yl)piperazin-1- —OCH(CH₃)₂ methylpyrazol- 3carbon atoms yl)C(O)O-]benzyl- 4-yl- (L-pyrrolidinyl) 367 1- R²/R³ =cyclic p-[(CH₃)₂NC(O)O-]benzyl- —OCH₂CH₂Oφ methylpyrazol-—CH₂—S—C(CH₃)₂— 4-yl- (L-5,5- dimethylthiazolidin-4- yl) 368 1- R²/R³ =cyclic 3-chloro-4-[(4-pyridin-2-yl)piperazin-1- —OH methylpyrazol-—CH₂—S—C(CH₃)₂— yl)C(O)O-]- 4-yl- (L-5,5- benzyl- dimethylthiazolidin-4-yl) 369 1- R²/R³ = cyclic 3 -chloro-4-[(4-pyridin-2-yl)piperazin-1-—OCH₂CH₃ methylpyrazol- —CH₂—S—C(CH₃)₂— yl)C(O)O-]- 4-yl- (L-5,5-benzyl- dimethylthiazolidin-4- yl) 370 1,5-dimethyl-3- R²/R³ = cyclicp-[4-[5-CF₃-pyridin-2-yl)piperazin-1 yl)- —OH chloropyrazol-—CH₂—S—C(CH₃)₂— C(O)O-]benzyl- 4-yl- (L-5,5- dimethylthiazolidin-4- yl)

In addition, Examples 319, 324, 325, 332, 333, 334, 335 and 349 in Table11 are exemplified as follows:

Example 319 Synthesis ofN-(Toluene-4-sulfonyl)-L-(4-hydroxy)prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine

The startingN-(toluene-4-sulfonyl)-L-(4-hydroxy)prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalaninetert-butyl ester (300 mg) was dissolved in formic acid (15 mL). Thereaction was stirred at room temperature for 72 hours. The solvent wasevaporated and the residue was purified using HPLC, reverse phase,20-80% CH₃CN/water. At a retention time of 10.75 minutes, 82 mg of thetitle compound was obtained, mp: 128-130° C.

Example 324 Synthesis ofN-(Toluene-4-sulfonyl)-L-(4-oxo)prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine

The startingN-(toluene-4-sulfonyl)-L-(4-oxo)prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine tert-butyl ester (130 mg) was dissolved informic acid (7 mL). The reaction was stirred at room temperature for 6hours. The solvent was evaporated in vacuo to give 150 mg of the desiredproduct, mp: 111-112° C.

Example 325 Synthesis ofN-(Toluene-4-sulfonyl)-L-(4-oxo)prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine

The startingN-(toluene-4-sulfonyl)-L-(4-oxo)prolyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalaninetert-butyl ester (150 mg) was dissolved in formic acid (7 mL). Thereaction was stirred at room temperature for 6 hours. The solvent wasevaporated in vacuo, and the residue was purified using HPLC, reversephase, 20-80% CH₃CH/water. The retention time was 10.34 minutes. Theproduct was freeze dried to yield 82 mg of the title compound, mp:99-101° C.

Example 332 Synthesis ofN-(Toluene-4-sulfonyl)-L-(4-methanesulfonyloxy)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The startingN-(toluene-4-sulfonyl)-L-(4-hydroxy)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester (300 mg) and methylsulfonyl chloride was dissolved inTHF (7 mL) at 0° C. in an ice bath. Triethylamine (0.21 mL) was added.The ice bath was removed after 10 minutes. The reaction mixture wasstirred at room temperature for 24 hours. Ethyl acetate (20 mL) asadded. The mixture was washed with citric acid (5%, 20 mL, 2×), andwashed with saturated NaHCO₃ solution (20 mL), then with brine. Thesolution was dried over MgSO₄. The solvent was evaporated, and theresidue was flushed on a silica gel column. The solvent was evaporatedin vacuo to give 300 mg of the desired product, mp: 73-74° C.

Example 333 Synthesis ofN-(4-Aminobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The startingN-(4-aminobenzenesulfonyl)-L-prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninemethyl ester (300 mg) and LiOH solution (2M, 0.6 mL) were added tomethanol (μmL). The reaction was stirred at room temperature for 7hours. The solvent was evaporated in vacuo, and the residue was purifiedusing HPLC, reverse phase, 20-80% CH₃CN/water. At a retention time of12.11 minutes, 27 mg of the desired product were obtained, mp: 130-132°C.

Example 334 Synthesis ofN-(4-Aminocarbonylbenzenesulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine

The startingN-(4-aminocarbonylbenzenesulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalaninemethyl ester (300 mg) and LiOH solution (2M, 0.5 mL) were added tomethanol (6 mL). The reaction was stirred at room temperature for 8hours. The solvent was evaporated in vacuo, and the residue purifiedusing HPLC, reverse phase, 20-80% CH₃CN/water. At a retention time of12.69 minutes, 20 mg of the desired product was obtained, mp: 123-125°C.

Example 335 Synthesis ofN-(4-Amidinobenzenesulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalanine

The startingN-(4-amidinobenzenesulfonyl)-L-prolyl-L-4-(thiomorpholin-4-ylcarbonyloxy)phenylalaninemethyl ester (300 mg) and LiOH solution (2M, 0.5 mL) were added tomethanol (6 mL). The reaction was stirred at room temperature for 8hours. The solvent was evaporated in vacuo, and the residue was purifiedusing HPLC, reverse phase, 20-80% CH₃CN/water. At a retention time of11.78 minutes, 25 mg of the desired product were obtained, mp: 123-125°C.

Example 349 Synthesis ofN-(Toluene-4-sulfonyl)-L-(4-methanesulfonyloxy)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The startingN-(toluene-4-sulfonyl)-L-(4-methanesulfonyloxy)prolyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester (200 mg) was dissolved in formic acid (5 mL). Thereaction mixture was stirred at room temperature for 6 hours. Thesolvent was evaporated in vacuo to provide the desired product (195 mg),mp: 83-84° C.

Example 371 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyi-4-(a-methylbenzyloxy)-L-phenylalanine

N-(Toluene-4-sulfonyl)-L-prolyl-L-tyrosine methyl ester (785 mg, 1.89mmol) was dissolved in DMF (20 mL) at room temperature. To this wasadded K₂CO₃ (1.1 eq, 281 mg) and 1-bromoethyl benzene (1.1 eq, 284 μL).The reaction was stirred for 12 hours at room temperature. Ethyl acetate(100 mL) was added, and the organic layer washed several times withbrine (5×50 mL). The organic layer was dried over MgSO₄. Upon filtrationand evaporation of the solvents under reduced pressure, an oil wasisolated. The crude material was purified by elution on silica gel(EtOAc/hexanes (1:4)). The desired material was isolated in 32% yield(330 mg, 0.6 mmol). The methyl ester (330 mg. 0.6 mmol) was thenconverted to the corresponding acid upon treatment with NaOH (1.1 eq, 27mg), in MeOH:H₂O (1:1) (15 mL), for 4 hours at room temperature. EtOAcwas added as well as water. The aqueous layer was collected andacidified with 1N HCl to pH 2.5, and reextracted with EtOAc. The organiclayer was dried over MgSO₄. Upon filtration and evaporation of thesolvents under reduced pressure, a foam was isolated in quantitativeyields.

NMR data was as follows:

¹H NMR (300 MHz, CDCl₃): δ=7.71 (bd, 2H), 7.34 (m, 7H), 7.20 (m, 1H),7.01 (m, 2H), 6.80 (d, 2H, J=8.37 Hz), 5.27 (m, 1H), 4.75 (m, 1H), 4.04(m, 1H), 3.23-2.93 (m, 4H), 2.42 (s, 3H), 1.85 (m, 1H), 1.60 (d, 3H,J=6.09 Hz), 1.36-1.26 (m, 3H).

¹³C NMR (75 MHz, CDCl₃): δ=174.74, 172.22, 157.53, 145.00, 143.77,133.42, 130.76, 130.58, 129.14, 128.60, 128.48, 127.94, 126.15, 116.57,76.39, 62.73, 53.90, 50.09, 37.09, 25.07, 24.52, 22.17.

Mass Spectroscopy: (FAB) 537 (M+H).

Example 372 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-4-(2-carboxyphenoxy)-L-phenylalanine

N-(Toluene-4-sulfonyl)-L-prolyl-L-tyrosine methyl ester (2.14 g, 5.16mmol) was added to a suspension of sodium hydride, 60% in oil (1.1 eq.,228 mg) in xylenes (50 mL) at 0° C. The reaction mixture was stirred for5 minutes and cuprous bromide dimethyl sulfide complex (1.4 eq., 1.48 g)was added. The reaction mixture was stirred at 23° C. for 0.5 hr. Tothis was added sodium 2-iodobenzoate (1.5 eq., 8.06 mmol), and thereaction mixture was refluxed for 12 hours. EtOAc (100 mL) was added,and the organic layer washed with NH₄Cl, 10% HCl, and brine, then driedover MgSO₄. The crude material was eluted on column chromatography(silica gel), with CHCl₃:MeOH (9:1), and isolated as an oil. The acidwas prepared by treatment with NaOH (1.1 eq), in MeOH:H₂O (1:1) for 4hours at room temperature. The diacid was isolated as a foam.

NMR data was as follows:

¹H NMR (300 MHz, CDCl₃): δ=7.71 (m, 2H), 7.29 (m, 4H), 7.19 (m, 4H),6.72 (m, 1H), 4.84 (m, 1H), 4.13 (m, 1H), 3.39 (m, 1H), 3.11 (m, 3H),2.43 (s, 3H), 1.89 (m, 1H), 1.48 (m, 3H).

¹³C NMR (75 MHz, CDCl₃): δ=172.67, 157.84, 155.89, 155.04, 145.17,133.61, 133.19, 133.08, 131.69, 131.02, 130.64, 128.42, 127.87, 124.24,120.04, 119.61, 116.12, 62.81, 50.31, 37.28, 30.69, 24.81, 22.15.

Mass Spectroscopy: (FAB) 553 (M+H).

Example 373 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-O-(benzyl)-L-tyrosine

N-(Toluene-4-sulfonyl)-L-Pro-OH was treated with (COCl)₂ and DMF inCH₂Cl₂ to give, after evaporation, N-(Toluene-4-sulfonyl)-L-Pro-Cl. Thisproduct was treated with L-Tyr(Bn)-OH and NaOH in THF and H₂O, to give,after acidification, extraction, drying with MgSO₄, and evaporation thetitle compound as a clear oil.

NMR data was as follows:

¹H NMR (DMSO-d₆, 300 MHz): δ=8.04 (d, J=8.2, 1H), 7.70 (d, J=8.1, 2H),7.42-7.21 (m, 6H), 7.15 (d, J=8.5, 2H), 6.90 (d, J=8.5, 2H), 5.04 (s,2H), 4.49-4.42 (m, 1H), 4.13-4.09 (m, 1H), 3.33-3.27 (m, 2H), 3.10-2.89(m, 3H), 2.38 (s, 3H), 1.60-1.35 (m, 4H).

¹³C NMR (DMSO-d₆, 75 MHz): δ=172.63, 170.81, 157.0, 143.6, 137.2, 133.8,130.3, 129.8, 129.4, 128.9, 128.4, 127.6, 125.3, 114.4, 69.1, 61.3,53.4, 49.0, 35.8, 30.4, 23.8, 21.0.

Mass Spectroscopy: (+FAB, 3-nitrobenzyl alcohol) 523 (M+H).

Example 374 Synthesis of N-(Toluene-4-sulfonyl)-L-prolyl-4-(1-H,2-oxo-3-methyltetrahydropyrimidin-1-yl)-L-phenylalanine

The title compound was prepared from the corresponding t-butyl esterusing the procedure described in Method 11.

NMR data was as follows:

¹H NMR (CDCl₃): δ=7.73 (d, 2H), 7.58 (d, 1H), 7.34 (d, 2H), 7.21 (d,2H), 7.17 (d, 2H), 4.79 (q, 1H), 4.15-4.11 (m, 1H), 3.68-3.63 (m, 2H),3.48-3.39 (m, 3H), 3.27 (dd, 1H), 3.17 (dd, 1H), 3.15-3.07 (m, 1H), 2.99(s, 3H), 2.43 (s, 3H), 2.16 -2.08 (m, 2H), 2.00-1.98 (m, 1H).

¹³C NMR (CDCl₃):δ=173.4, 172.2, 164.2, 156.4, 144.4, 142.5, 134.1,133.0, 130.2, 130.0, 127.9, 126.2, 62.1, 53.4, 49.5, 48.9, 47.9, 36.5,35.9, 30.2, 24.2, 22.0, 21.4.

Example 375 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-4-(2-methoxyphenyl)-L-phenylalanine

The title compound was prepared from the corresponding t-butyl esterusing the procedure described in Method 11.

NMR data was as follows:

¹H NMR (CD₃OD):δ=7.70 (m, 2H), 7.36 (m, 4H), 7.22 (m, 4H), 6.98 (m, 2H),4.75 (m, 1H), 4.10 (m, 1H), 3.71 (s, 3H), 3.29 (m, 2H), 3.11 (m, 2H),2.39 (s, 3H), 1.75 (m, 1H), 1.53 (m, 3H).

¹³CNMR(CD₃OD): δ=174.4, 174.2, 158.1, 145.9, 138.9, 136.7, 135.1, 131.2,130.9, 130.8, 130.2, 129.9, 129.1, 122.0, 112.6, 63.3, 55.9, 54.6, 50.5,37.9, 31.5, 25.2, 21.4.

Example 376 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-4-(2-methoxyphenyl)-L-phenylalanine

The title compound was prepared from the corresponding t-butyl esterusing the procedure described in Method 11.

NMR data was as follows:

¹H NMR (CDCl₃): δ 8.41 (d, 1H), 8.21 (s, 1H), 8.03 (d, 1H), 7.98 (s,1H), 7.74 (d, 2H), 7.39 (d, 1H), 7.33 (d, 2H), 4.72-4.68 (m, 1H),4.17-4.13 (m, 1H), 3.54-3.34 (m, 3H), 3.20-3.12 (m, 1H), 2.82 (s, 6H)m2.43 (s, 3H), 2.09-2.04 (m, 1H), 1.79-1.59 (m, 3H).

¹³C NMR (CDCl₃): δ=173.7, 171.8, 154.5, 147.2, 144.4, 137.8, 135.5,133.2, 130.1, 127.9, 126.4, 62.2, 53.0, 49.5, 38.5, 36.0, 30.3, 24.4,21.4.

Example 377 Synthesis ofN-(Toluene-4-sulfonyl)-L-prolyl-4-(2,4,5-trioxo-3-(3-chlorophenyl)-tetrahydroimidazol-1-yl)-L-phenylalanine benzyl ester

The compound was prepared by treatment ofN-(toluene-4-sulfonyl)-L-prolyl-4-[(3- chlorophenylureido)-tetrahydroimidazol-1 -yl]-L-phenylalanine isopropyl ester withoxalyl chloride in methylene chloride. The crude product was purified byflash chromatography (silica, 3:2 Hex: EtOAc) to afford a white solid.(0.410 g, 50%).

MS ((+)ESI, m/z (%) 746 (100[M+H]+) (746/748 C1)

Example 378 Synthesis ofN-(Phenyl-sulfonyl)-D-prolyl-L-4-(2,6-dimethoxyphenyl)phenylalanine

The title compound was prepared by coupling of2,6-dimethoxyphenylboronic acid and 4′-iodophenylalanine derivates toprovide dimethoxybiphenylalanines such as the title compound followingprocedures outlined in Hagmann et al., Bioorganic & Medicinal ChemistryLetters, 2001; 11 (20): 2709-2713; Kamenecka et al., Bioorganic &Medicinal Chemistry Letters, 2002; 12(16): 2205-2208; and Doherty etal., Bioorganic & Medicinal Chemistry Letters, 2003; 13(11): 1891-1895.

Example 379 Synthesis ofN-(3,5-dichlorophenyl-sulfonyl)-D-prolyl-L-4-[4-(methylcarbonylaminobutyl)-2,5-Dioxo-imidazolidin-1-yl]phenylalanine

The title compound was prepared following procedures outlined in WO01/54690.

Example 380 Synthesis ofN-(2,6-dichlorophenyl-carbonyl)-L-4-(2,6-dimethoxyphenyl)phenylalanine

The title compound was prepared by coupling of2,6-dimethoxyphenylboronic acid and 4′-iodophenylalanine derivates toprovide dimethoxybiphenylalanines such as the title compound followingprocedures outlined in WO 99/36393 and Sircar et al., Bioorganic &Medicinal Chemistry, 2002; 10(6): 2051-2066.

Example 381 Synthesis ofN-[N-(3-pyridinesulfonyl)-L-3,3-dimethyl-4-thiaprolyl]-O-[1-methylpiperazin-4-ylcarbonyl]-L-tyrosineisopropyl ester

3-Pyridinesulfonyl chloride

The free base of the title compound may be prepared from3-pyridinesulfonic acid (Aldrich) by a published procedure: Corey etal., J. Org. Chem. 1989, 54(2): 389. Alternatively, the hydrochloride ofthe title compound may be prepared from 3-pyridinesulfonic acid(Aldrich) by published procedures: Crowell et al., J. Med. Chem. 1989,32(11): 2436; Karaman et al., J. Am. Chem. Soc. 1992, 114(12): 4889.

L-3,3 -Dimethyl-4-thiaproline

The title compound may be prepared from L-penicillamine (Aldrich) bypublished procedures: Samanen et al., J. Med. Chem. 1989, 32(2): 466;Nagasawa et al., J. Med. Chem. 1984, 27(5): 591.

N-(3 -pyridinesulfonyl)-L-3,3 -dimethyl-4-thiaproline

A pH=7.4 buffer was prepared by dissolving disodium hydrogen phosphate(43.2 g, 0.304 mol) and potassium dihydrogen phosphate (11.8 g, 0.0870mol) in H₂O to give a volume of 1.0 L. To a 0° C. solution ofL-3,3-dimethyl-4-thiaproline (25.4 g, 0.157 mol) in 700 mL pH=7.4 bufferwas added with stirring a solution of 3-pyridinesulfonyl chloride (28.0g, 0.157 mol) in 300 mL CH₂Cl₂. The mixture was stirred for 24 h whilewarming to room temperature, and was acidified to pH=2 by addition of 3M H₂SO₄, precipitating a yellow solid. The yellow solid was isolated byfiltration of both phases, and the CH₂Cl₂ layer was separated andevaporated to afford additional yellow solid. The combined yellow solidswere stirred in 700 mL H₂O for 1 h, to dissolve associated inorganicsalts, and isolated again by filtration. The two aqueous layers werecombined and extracted with EtOAc (3×500 mL). The EtOAc layers werewashed with brine, treated with sodium sulfate, filtered, and evaporatedto afford additional yellow solid. All aliquots of yellow solid werecombined to afford 36.1 g (76%)N-(3-pyridinesulfonyl)-L-3,3-dimethyl-4-thiaproline.

N-[N-(3-pyridinesulfonyl)-L-3,3-dimethyl-4-thiaprolyl]-L-trosineisopropyl ester

1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (4.83 g,0.0253 mol) was added to a 0° C. solution ofN-(3-pyridinesulfonyl)-L-3,3-dimethyl-4-thiaproline (6.37 g, 0.0211mol), L-tyrosine isopropyl ester hydrochloride (5.48 g, 0.0211 mol),1-hydroxybenzotriazole (5.69 g, 0.0421 mol), and 4-methylmorpholine(2.32 mL, 2.13 g, 0.0211 mol) dissolved in 125 mL DMF. The mixture wasstirred for 16 h while warming to room temperature, and 200 mL EtOAc and200 mL H₂O were added. The mixture was shaken, and the aqueous layer wasseparated, and the organic layer was washed with 0.2 M citric acid(2×100 mL), H₂O (2×100 mL), sat. NaHCO₃ (2×100 mL), H₂O (2×100 mL), andbrine (2×100 mL). The organic layer was treated with sodium sulfate,filtered, and evaporated to afford 9.40 g (86%)N-[N-(3-pyridinesulfonyl)-L-3,3-dimethyl-4-thiaprolyl]-L-tyrosineisopropyl ester as a yellow foam. ¹H NMR (CDCl₃, 300 MHz) δ 9.08 (bs,1H), 8.86 (bs, 1H), 8.16 (dt, J_(d)=8.1 Hz, J₂=2.0 Hz, 1H), 7.51 (dd,J=8.0 Hz, J=4.6), 7.07 (d, J=8.1 Hz, 2H), 6.87 (d, J=8.1 Hz, 1H), 6.74(d, J=8.1 Hz, 2H), 5.96 (bs, 1H), 5.06 (sept, J=6.3, 1H), 4.83 (dt,J_(d)=6.0 Hz, J_(t)=7.8 Hz, 1H), 4.57 (d, J=9.3 Hz, 1H), 4.46 (d, J=9.3Hz, 1H), 3.91 (s, 1H), 3.09 (dd, J=14.1 Hz, J=5.4 Hz, 1H), 2.98 (dd,J=14.1 Hz, J=7.5 Hz, 1H), 1.25 (t, J=6.6 Hz, 6H), 1.18 (s, 3H), 1.13 (s,3H). ¹³C NMR (CDCl₃, 75 MHz) δ 170.5, 168.0, 155.2, 154.2, 148.6, 135.9,130.7, 127.6, 124.1, 115.5, 105.5, 73.7, 69.7, 54.7, 53.4, 50.5, 37.5,29.2, 23.7, 21.62, 21.55.

N-[N-(3-pyridinesulfonyl)-L-3,3-dimethyl-4-thiaprolyl]-O-[1-methylpiperazin-4-ylcarbonyl]-L-tyrosineisopropyl ester

N-[N-(3-pyridinesulfonyl)-L-3,3-dimethyl-4-thiaprolyl]-L-tyrosineisopropyl ester (1.51 g, 2.89 mmol) and 4-nitrophenyl chloroformate(0.58 g, 2.89 mmol) were dissolved in 40 mL CH₂Cl₂, and the solution wasstirred for 15 min while cooling in a 15° C. slurry of 41 H₂O/EtOH anddry ice. To the solution was added Et₃N (1.00 mL, 0.73 g, 7.23 mol) withstirring over 2 min, and the solution was stirred for 1 h at −15° C. Tothe resulting suspension was added 1-methylpiperazine (0.32 mL, 0.289 g,2.89 rnmol) with stirring over 1 min, and the mixture was stirred for 16h while warming to room temperature. The mixture was diluted with 40 mLhexanes, and washed with 10% (w/v) K₂CO₃ (4×50 mL) until no yellow color(4-nitrophenol) was seen in the aqueous layer. The organic layer waswashed with brine (75 mL), treated with sodium sulfate, filtered, andevaporated to give a light yellow residue. The residue was purified bychromatography on silica gel using 70:25:5 CH₂Cl₂/EtOAc/EtOH to afford1.53 g (84 %) N-[N-(3-pyridinesulfonyl)-L-3,3-dimethyl-4-thiaprolyl]-O-[1-methylpiperazin-4-ylcarbonyl]-L-tyrosine isopropylester as a colorless foam. ¹H NMR (CDCl₃, 300 MHz) δ 9.09 (d, J=2.1 Hz,1H), 8.87 (dd, J=4.9 Hz, J=1.6 Hz, 1H), 8.16 (dt, J_(d)=8.4 Hz,J_(t)=2.0 Hz, 1H), 7.51 (dd, J=8.2 Hz, J=4.9 Hz, 1H), 7.21 (d, J=8.4 Hz,2H), 7.02 (d, J=8.4 Hz, 2H), 6.89 (d, J=7.8 Hz, 1H), 5.05 (sept, J=6.4Hz, 1H), 4.84 (q, J=7.0 Hz, 1H), 6.59 (d, J=9.9 Hz, 1H), 4.47 (d, J=9.9Hz, 1H), 3.90 (s, 1H), 3.67 (bs, 2H), 3.58 (bs, 2H), 3.18-3.03 (m, 2H),2.45 (t, J=10.2 Hz, 4H), 2.34 (s, 3H), 1.26 (d, J=6.0 Hz, 3H), 1.23 (d,J=6.6 Hz, 3H), 1.20 (s, 3H), 1.17 (s, 3H). ¹³C NMR (CDCl₃, 75 MHz) δ170.4, 167.8, 154.3, 153.7, 150.6, 148.7, 135.8, 133.1, 133.0, 130.4,133.0, 121.8, 73.7, 69.7, 54.8, 54.6, 54.5, 50.5, 46.1, 44.3, 43.8,37.6, 29.1, 23.8, 21.6, 21.5.

Synthesis of Compounds of Formulae III-IX

The following Methods may be used to prepare the compounds of FormulaeIII-IX.

Method A Methyl Ester Preparation Procedure

Amino acid methyl esters can be prepared using the method of Brenner andHuber Helv. Chim. Acta 1953, 36, 1109.

Method B BOP Coupling Procedure

The desired dipeptide ester was prepared by the reaction of a carboxylicacid (1 equivalent) with the appropriate amino acid ester or amino acidester hydrochloride (1 equivalent),benzotriazol-1-yloxy-tris(dimethylamino)phosphonium hexafluorophosphate[BOP] (2.0 equivalent), methylamine (1.1 equivalent), and DMF. Thereaction mixture was stirred at room temperature overnight. The crudeproduct is purified flash chromatography to afford the dipeptide ester.

Method C Hydrogenation Procedure I

Hydrogenation was performed using 10% palladium on carbon (10% byweight) in methanol at 30 psi overnight. The mixture was filteredthrough a pad of Celite and the filtrate concentrated to yield thedesired compound.

Method D Hydrolysis Procedure I

To a chilled (0° C.) THF/H₂O solution (2:1, 5-10 mL) of the appropriateester was added LiOH (or NaOH) (0.95 equivalents). The temperature wasmaintained at 0° C. and the reaction was complete in 1-3 hours. Thereaction mixture was extracted with ethyl acetate and the aqueous phasewas lyophilized resulting in the desired carboxylate salt.

Method E Ester Hydrolysis Procedure II

To a chilled (0° C.) THF/H₂O solution (2:1, 5-10 mL) of the appropriateester was added LiOH (1.1 equivalents). The temperature was maintainedat 0° C. and the reaction was complete in 1-3 hours. The reactionmixture was concentrated and the residue was taken up into H₂O and thepH adjusted to 2-3 with aqueous HCl. The product was extracted withethyl acetate and the combined organic phase was washed with brine,dried over MgSO₄, filtered and concentrated to yield the desired acid.

Method F Ester Hydrolysis Procedure III

The appropriate ester was dissolved in dioxane/H₂O (1:1) and 0.9equivalents of 0.5 N NaOH was added. The reaction was stirred for 3-16hours and then concentrated. The resulting residue was dissolved in H₂Oand extracted with ethyl acetate. The aqueous phase was lyophilized toyield the desired carboxylate sodium salt.

Method G

BOC Removal Procedure

Anhydrous hydrochloride (HCl) gas was bubbled through a methanolicsolution of the appropriate Boc-amino acid ester at 0° C. for 15 minutesand the reaction mixture was stirred for three hours. The solution wasconcentrated to a syrup and dissolved in Et₂O and reconcentrated. Thisprocedure was repeated and the resulting solid was placed under highvacuum overnight.

Method H tert-Butyl Ester Hydrolysis Procedure I

The tert-butyl ester was dissolved in CH₂Cl₂ and treated with TFA. Thereaction was complete in 1-3 hr at which time the reaction mixture wasconcentrated and the residue dissolved in H₂O and lyophilized to yieldthe desired acid.

Method I EDC Coupling Procedure I

To a CH₂Cl₂ solution (5-20 mL) of a carboxylic acid (1 equivalent), theappropriate amino acid ester hydrochloride (1 equivalent),N-methylmorpholine (1.1-2.2 equivalents) and 1-hydroxybenzotriazole (2equivalents) were mixed, placed in an ice bath and1-(3-dimethylaminopropyl)-3-ethyl carbodiimide (1.1 equivalents) added.The reaction was allowed to rise to room temperature and stirredovernight. The reaction mixture was poured into H₂O and the organicphase was washed with sat. NaHCO₃, brine, dried (MgSO₄ or Na₂SO₄),filtered and concentrated. The crude product was purified by columnchromatography.

Method J EDC Coupling Procedure II

To a DMF solution (5-20 mL) of a carboxylic acid (1 equivalent), theappropriated amino acid ester hydrochloride (I equivalent), Et₃N (1.1equivalents) and 1-hydroxybenzotriazole (2 equivalents) were mixed,placed in an ice bath and 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide(1.1 equivalents) added. The reaction was allowed to rise to roomtemperature and stirred overnight. The reaction mixture was partitionedbetween EtOAc and H₂O and the organic phase washed with 0.2 N citricacid, H₂O, sat. NaHCO₃, brine, dried (MgSO₄ or Na₂SO₄) filtered andconcentrated. The crude product was purified by column chromatography orpreparative TLC.

Method K tert-Butyl Ester Hydrolysis Procedure II

The tert-butyl ester was dissolved in CH₂Cl₂ (5 mL) and treated with TFA(5 mL). The reaction was complete in 1-3 hours at which time thereaction mixture was concentrated and the residue dissolved in H₂O andconcentrated. The residue was redissolved in H₂O and lyophilized toyield the desired product.

Method L Carbamate Formation Procedure I

Into a reaction vial were combined 15.2 mmol, 1.0 eq. of the startinghydroxy compound (typically a tyrosine derivative) and 1.86 g (15.2mmol, 1.0 eq) DMAP. Methylene chloride (50 mL), triethylamine (2.12 mL,1.54 g, 15.2 mmol, 1.0 eq), and dimethylcarbamyl chloride (1.68 mL, 1.96g, 18.2 mmol, 1.2 eq) were then added. The vial was capped tightly, andthe reaction solution swirled to obtain a homogeneous solution. Thereaction solution was then heated to 40° C. After 48 h, TLC of theresulting colorless solution indicated complete conversion. The work-upof the reaction solution was as follows: 50 mL EtOAc and 50 mL hexaneswas added to the reaction mixture, and the resulting mixture was washedwith 0.5 M citric acid (3×50 mL), water (2×50 mL), 10% K₂CO₃ (2×50 mL),and sat. NaCl (1×50 mL); dried with MgSO₄, filtered and evaporated toafford the desired compound.

Method M Carbamate Formation Procedure II

Into a reaction vial were combined 84.34 mmol (1.0 eq) of the startinghydroxy compound (typically a tyrosine derivative) and 17.0 g (84.34mmol, 1.0 eq) 4-nitrophenyl chloroformate. Methylene chloride (700 mL)was added and the vial was capped with a septum. A nitrogen line wasattached and the vial was immersed in a 4:1 water/ethanol dry ice slurrywith stirring to cool to −15° C. Triethylamine (29.38 mL, 21.33 g,210.81 mmol, 2.5 eq) was added over five minutes with stirring and thestirring was continued at −10 to −15° C. for 1 h. N-Methyl piperazine(9.35 mL, 8.45 g, 84.34 mmol, 1.0 eq) was added over three minutes withstirring and stirring was continued overnight while warming to room-temperature. The reaction mixture was diluted with 700 mL hexanes andthe resulting mixture was washed repeatedly with 10% K₂CO₃, until noyellow color (from 4-nitrophenol) is observed in the aqueous layer. Themixture was then washed with sat. NaCl, dried over anhydrous MgSO₄,filtered and evaporated. The residue was dissolved in 500 mL of ethanoland evaporated to remove triethylamine. The residue was again dissolvedin 500 mL of ethanol and evaporated to remove triethylamine. The residuewas then dissolved in 400 mL of ethanol and 600 mL of water was addedwith stirring to precipitate a solid or oil. If an oil if formed, theoil is stirred vigorously to induce it to solidify. The solid is thenisolated by filtration. Dissolution, precipitation, and filtration arerepeated once and the resulting solid is rinsed with water to removetraces of yellow color. The solid is then subjected to high vacuum untilthe mass remains constant thereby affording the desired carbamyloxycompound.

Method N Preparation of 5-Iodo-4(3H)-pyrimidinone

The procedure of Sakamoto et. al. (Chem. Pharm. Bull. 1986, 34(7),2719-2724) was used to convert 4(3H)-pyrimidinone into5-iodo-4(3H)-pyrimidinone, which was of sufficient purity for conversionto 4-chloro-5-iodopyrimidine.

Method O Preparation of 4-Chloro-5-iodopyrimidine

5-Iodo-4(3H)-pyrimidinone (1 eq.) was suspended in toluene to which wasadded POCl₃ (2.0 eq.). The reaction mixture was heated to reflux for 3hours, and then cooled and concentrated. The residue was suspended inwater, adjusted to pH=7 by addition of 4N sodium hydroxide, andextracted with ethyl acetate. The organic extracts were washed withbrine, dried (MgSO₄), filtered and stripped to give a red oil. The crudeproduct was dissolved in methanol and silica gel was added. Followingconcentration, the coated silica gel was loaded onto a plug of silicagel and elution with ethyl acetate/hexanes yielded the title compound.

Method P Preparation ofN-(5-Iodopyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenvlalaninetert-Butyl Ester

A solution 4-chloro-5-iodopyrimidine (1.0 eq.),L-4-(N,N-dimethylcarbamyloxy)-phenylalanine tert-butyl ester (1.0 eq),and N,N-diisoproylethyl amine (2.0 eq) in tetrahydrofuran was heated atreflux for 16 hours. The reaction mixture was then cooled and dilutedwith water and ethyl acetate. The organic phase was washed with 0.2 Ncitric acid, water, saturated NaHCO₃, brine, dried (MgSO₄), filtered andconcentrated. The residue was purified by silica gel chromatographyusing ethyl acetate/hexanes to afford the title compound.

Method Q Suzuki Coupling Procedure I

To an ethyleneglycol dimethyl ether solution oftetrakis(triphenylphosphine)palladium (0.04 eq) was addedN-(5-iodopyrimidin-4-yl)-L4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester (1.0 eq.). After stirring for approximately ten minutesa boronic acid or ester (1.2 eq) and 2M Na₂CO₃ (2.0 eq) were added, andthe reaction flask was evacuated and then flushed with nitrogen gas. Thereaction was heated at reflux from three to sixteen hours. The reactionmixture was then cooled, diluted with water and ethyl acetate, and theorganic phase was washed with 0.2 N citric acid, water, saturatedNaHCO₃, brine, dried (MgSO₄), filtered and concentrated. Alternatively,the cooled reaction mixture was diluted with ethyl acetate and washedwith water, saturated NaHCO₃, dried (MgSO₄), filtered and concentrated.Either column chromatography or preparative thin layer chromatography onsilica gel using ethyl acetate/hexanes afforded the desired product.

Method R Suzuki Coupling Procedure II

To a dimethylformamide solution oftetrakis(triphenylphosphine)-palladium (0.02-0.05 eq) was addedN-(5-iodopyrimidin-4-yl)-L-4-(N,N dimethylcarbamyloxy)phenylalaninetert-butyl ester (1.0 eq.). After stirring for approximately tenminutes, the boronic acid (1.1-4.0 eq) and K₃PO₄ (1.5-2.0 eq) wereadded, and the reaction was heated at 100° C. for three to sixteenhours. The reaction mixture was then cooled, diluted with water andethyl acetate, and the organic phase was washed with 0.2 N citric acid,water, saturated NaHCO₃, brine, dried (MgSO₄), filtered andconcentrated. Either column chromatography or preparative thin layerchromatography on silica gel using ethyl acetate/hexanes afforded thedesired product

Method S Suzuki Coupling Procedure III

An ethyleneglycol dimethyl ether/2M Na₂CO₃ (1:1 by volume) solution oftetrakis(triphenylphosphine)palladium (0.04 eq),N-(5-iodopyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester (1.0 eq.), the boronic acid (1.1 eq) and lithiumchloride (3.0 eq ) was heated to reflux for approximately six hours. Thecooled reaction mixture was diluted with ethyl acetate and washed withwater, brine, dried (MgSO₄), filtered and concentrated. The residue waspurified by silica gel column chromatography using ethyl acetate/hexanesto afford the desired product.

Method T Suzuki Coupling Procedure IV

An ethyleneglycol dimethyl ether/2M Na₂CO₃, (1:1 by volume) solution oftetrakis(triphenylphosphine)palladium (0.05 eq),N-(5-iodopyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester (1.0 eq.), the boronic acid (1.5 eq) andtri-o-tolylphosphine (0.1 eq) was heated to reflux for approximatelythree hours. The cooled reaction mixture was diluted with ethyl acetateand water and washed with water, brine, dried (MgSO₄), filtered andconcentrated. The residue was purified by preparative thin layerchromatography on silica gel using ethyl acetate/hexanes to afford thedesired product.

Method U Heck Reaction Procedure I

A dimethylformamide solution ofN-(5-iodopyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester (1.0 eq.), N,N-dimethylacrylamide (2.0 eq), andtriethylamine (6.0 eq) was degassed with nitrogen and thendichlorobis-(triphenylphosphine)palladium was added. The reaction waswarmed to 90° C. under a stream of nitrogen for 16 hours. The cooledreaction mixture was diluted with ethyl acetate and water and washedwith water, brine, dried (MgSO₄), filtered and concentrated. The residuewas purified by column chromatography on silica gel using ethylacetate/hexanes followed by preparative thin layer chromatography onsilica gel using ethyl acetate/hexanes to afford the desired product.

Method V Hydrogenation Procedure II

N-(5-(2-N,N-dimethylcarbamylethyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester was dissolved in ethanol to which was added 10%palladium on carbon. The reaction mixture was hydrogenated at 35 psihydrogen for approximately five hours. The reaction mixture was filteredthrough a pad of Celite, and the filtrate was concentrated. The residuewas purified by preparative thin layer chromatography on silica gelusing methanol/dichloromethane to afford the desired product.

Method W Heck Reaction Procedure II

To a tetrahydrofuran solution ofN-(5-iodopyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester (1.0 eq) dichlorobis(triphenylphosphine)palladium,triethylamine (0.05 eq) and triphenylphosphine (0.025 eq) was addedphenylacetylene (1.5 eq) and triethylamine (1.5 eq). After twentyminutes, copper (I) iodide (0.012 eq) was added, and the resultingmixture was stirred overnight at room temperature. The reaction mixturewas then diluted with ethyl acetate and water and washed with 0.2 Ncitric acid, water, saturated NaHCO₃, brine, dried (MgSO₄), filtered andconcentrated. The residue was chromatographed on a silica gel columnusing ethyl acetate/hexanes. ¹H NMR analysis showed that the desiredproduct to be contaminated with the iodopyrimidine starting material.However, the product was used without further purification.

Method X Hydrogenation Procedure III

Crude N-(5-(2-phenylethynyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine tert-butyl ester was dissolved inethanol to which was added 10% palladium on carbon and sodium acetate(3.0 eq). The reaction mixture was hydrogenated at 40 psi hydrogen forapproximately three hours, then filtered through a pad of Celite, andthe filtrate concentrated. The residue was washed with 0.2 N citricacid, water, saturated NaHCO₃, brine, dried (MgSO₄), filtered andconcentrated. Silica gel column chromatography using ethylacetate/hexanes yielded the desired product.

Method Y Preparation ofN-(6-Chloropyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

A solution 4,6-dichloropyrimidine (1.2 eq),L-4-(N,N-dimethylcarbamyloxy)-phenylalanine tert-butyl ester (1.0 eq),and triethylamine (1.05 eq) in ethanol was heated at reflux for 16hours. The reaction mixture was cooled and concentrated, and the residuewas taken-up in water and ethyl acetate. The organic phase was washedwith 0.2 N citric acid, water, saturated NaHCO₃, brine, dried (MgSO₄),filtered and concentrated. The residue was purified by silica gelchromatography using ethyl acetate/hexanes to afford the title compound.

Method Z Suzuki Coupling Procedure V

An ethyleneglycol dimethyl ether solution oftetrakis(triphenylphosphine)palladium (0.12 eq),N-(6-chloropyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester (1.0 eq.) and triphenylphosphine (0.05 eq) was stirredfor approximately ten minutes. The boronic acid or ester (1.2-2.5 eq)and 2M Na₂CO₃ (2.0 eq) were added, and the reaction was heated at 90° C.for 16 to 72 hours. The reaction mixture was cooled and concentrated,and the residue was taken up in water and ethyl acetate. The organicphase was washed with 0.2 N citric acid, water, saturated NaHCO₃, brine,dried (MgSO₄), filtered and concentrated. The residue was purified bypreparative thin layer chromatography on silica gel using ethylacetate/hexanes to afford the desired product.

Method AA Preparation ofN-(6-(N-Alkylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloy)Phenylalaninetert-Butyl Ester

A mixture ofN-(6-chloropyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester (1.0 eq) and an alkylamine (10.0 eq) was heated in asealed tube at 120° C. for 16 hours. The reaction mixture was cooled anddiluted with ethyl acetate. The organic portion was washed with 0.2 Ncitric acid, water, saturated NaHCO₃, brine, dried (MgSO₄), filtered andconcentrated. The residue was purified by silica gel chromatographyusing ethyl acetate/hexanes to afford the desired compound.

Method BB Preparation of 4-N-Alkylamino-5-bromo-2-chloropyrimidine

A methanol solution of 5-bromo-2,4-dichloropyrimidine (1.0 eq), thealkylamine (1.05 eq , typicallyL-4-(N,N-dimethylcarbamyloxy)phenylalanine tert-butyl ester), andN,N-diisoproylethylamine (5.0 eq) was heated to 40° C. for 16 hours. Thereaction mixture was then concentrated, and the residue was taken up inethyl acetate. The organic portion was washed with 0.2 N citric acid,water, saturated NaHCO₃, brine, dried (MgSO₄), filtered andconcentrated. The crude material was purified by silica gelchromatography using ethyl acetate/hexanes to afford the desiredcompound.

Method CC Preparation of 4-N-Alkylamino-5-bromo-2-N-alkylaminopyrimidine

An isopropanol solution of the 4-N-alkylamino-5-bromo-2-chloropyrimidine(1.0 eq) and an alkylamine (5.0 eq) was heated in sealed tube at 130° C.for 3-5 hours. The reaction mixture was then cooled and washed with 0.2N citric acid, water, saturated NaHCO₃, brine, dried (MgSO₄), filteredand concentrated. The crude material was purified by silica gelchromatography using ethyl acetate/hexanes to afford the desiredcompound.

Method DD 4-N-Alkylamino-5-bromo-2-N-alkylaminopyrimidine SuzukiCoupling Procedure

To an ethyleneglycol dimethyl ether solution oftetrakis(triphenylphosphine)palladium (0.04 eq) was added an4-N-alkylamino-5-bromo-2-N- alkylaminopyrimidine (1.0 eq.). Afterstirring for approximately ten minutes, the boronic acid or ester (1.2eq) and 2M Na₂CO₃ (2.0 eq) was added, and the reaction flask wasevacuated and then flushed with nitrogen gas. The reaction was heated atreflux for three to four hours. The reaction mixture was then cooled anddiluted with water and ethyl acetate, and the organic phase was washedwith 0.2 N citric acid, water, saturated NaHCO₃, brine, dried (MgSO₄),filtered and concentrated. The residue was purified by either silica gelcolumn or preparative thin layer chromatography using ethylacetate/hexanes to afford the desired product.

Method EE Preparation of N-tert-Butoxyvcarbonyl-4-Iodo-L-phenyalanineMethyl Ester

The title compound was prepared from 4-iodo-L-phenylalanine by standardconditions described by Bodanszky and Bodanszky in The Practiceofpeptide Synthesis; Springer-Verlag: Berlin, 1984.

Method FF Preparation ofN-tert-Butoxycarbonyl-4-(2,6-dimethoxyphenyl)-L-phenyalanine MethylEster

To a dimethylformamide solution of tetrakis(triphenylphosphine)palladium(0.02 - 0.05 eq) was addedN-tert-butoxycarbonyl-4-(2,6-dimethoxyphenyl)-L-phenyalanine methylester (1.0 eq.). After stirring for approximately ten minutes,2,6-dimethoxyphenyl boronic acid (1.1 eq) and K₃PO₄ (2.0 eq) were added,and the reaction was heated at 100° C. for sixteen hours. The reactionmixture was then cooled, diluted with water and ethyl acetate, and theorganic phase was washed with 0.2 N citric acid, water, saturatedNaHCO₃, brine, dried (MgSO₄), filtered and concentrated. Columnchromatography on silica gel using ethyl acetate/hexanes afforded thedesired product.

Method GG Preparation of 4-(2,6-Dimethoxyphenyl)-L-phenyalanine MethylEster Trifluoroacetic Acid Salt

A methylene chloride solution ofN-tert-butoxycarbonyl-4-(2,6-dimethoxyphenyl)-L-phenyalanine methylester was treated with trifluoroacetic acid for six hours at roomtemperature. Concentration of the volatiles yielded the title compound.

Method HH tert-Butyl Ester Cleavage Procedure III

A methylene chloride solution of the appropriate tert-butyl ester wastreated with trifluoroacetic acid at room temperature. After 2-3 hoursthe volatiles were evaporated, and the residue was treated again withmethylene chloride and trifluoroacetic acid. After 2-3 hours thevolatiles were evaporated again to yield the desired compound.

Method II Preparation ofN-(5-Allylpyrimidin-4-yl)-L-4-(N,N-dimethyl-carbamyloxy)phenylalaninetert-Butyl Ester

N-(5-iodo-pyrimidin-4-yl)-L-4-(N,N-dimethyl-carbamyloxy)phenylalaninetert-butyl ester (1.0 eq) was dissolved in dry DMF, withallyltributylstannane (1.1 eq), bis(triphenylphosphine)palladiumdichloride (0.03 eq) and LiCl (3.Oeq). The reaction mixture was flushedunder nitrogen, and heated to 90° C. for 2 hours. EtOAc was added, andthe organic layer was washed with water and brine, and dried over MgSO₄.After filtration and evaporation of the solvent under reduced pressure,the crude material was purified by column chromatography (silica gel)eluting with EtOAc/hexanes 1:3. The title material was isolated in goodyields.

Method JJ Preparation ofN-[5-propylpyrimidin-4-yll-L-4-(N,N-dimethyl-carbamyloxy)phenylalaninetert-Butyl Ester

N-(5-Allylpyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester was dissolved in methanol and treated with a catalyticamount of 10% palladium on carbon. The mixture was shaken under 10 psihydrogen gas for 3 hours. Upon filtration though a pad of Celite, andevaporation of the solvent under reduced pressure, the desired materialwas isolated as a foam.

Method KK Preparation ofN-(5-propylpyrimidin-4-yl)-L-4-(N,N-dimethyl-carbamyloxyphenylalanine

N-(5-Propylpyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester was treated with neat trifluoroacetic acid, and themixture was stirred for 5 h at room temperature. Upon evaporation of thesolvent under reduced pressure, the desired material was isolated as afoam.

Method LL Preparation of Dimethyl 2-Alkylmalonate

To a suspension of sodium hydride 60% dispersion in mineral oil (1.1 eq)in anhydrous THF was added slowly with stirring dimethyl malonate (1.1eq), causing the evolution of gas. To the resulting solution was added abromoalkane, iodoalkane, or trifluoromethanesulfonyloxyalkane (1.0 eq),and the mixture was heated to 50° C. for 48 h, at which point TLCindicated consumption of the bromoalkane, iodoalkane, ortrifluoromethanesulfonyloxyalkane. The mixture was diluted with diethylether and washed with 70% saturated sodium chloride. The organicextracts were treated with anhydrous magnesium sulfate, filtered, andevaporated to afford a dimethyl 2-alkylmalonate of sufficient purity forimmediate conversion to a 5-alkyl-4,6-dihydroxypyrimidine.

Method MM Preparation of Diethyl 2-Alkylidenylmalonate

Procedure B (p. 2759) of Houve and Winberg (J. Org. Chem. 1980, 45(14),2754-2763) was employed to react diethyl malonate with a ketone or analdehyde to afford a diethyl 2-alkylidenylmalonate of sufficient purityfor immediate conversion to a diethyl 2-alkylmalonate.

Method NN Preparation of Diethyl 2-Alkylmalonate

A diethyl 2-alkylidenylmalonate and an equal mass 10% palladium oncarbon were suspended in ethanol. The mixture was shaken under 55 psihydrogen gas for 24 h, at which point TLC indicated consumption of thediethyl 2-alkylidenylmalonate. The mixture was filtered through Celiteand evaporated to afford a diethyl 2-alkylmalonate of sufficient purityfor immediate conversion to a 5-alkyl-4,6-dihydroxypyrimidine.

Method OO Preparation of 5-Alkyl-4,6-dihydroxypyrimidine

To a diethyl 2 alkylmalonate or a dimethyl 2-alkylmalonate (1.0 eq) wasadded formamidine acetate (1.0 eq) and 25% sodium methoxide in methanol(3.3 eq). The resulting slurry was stirred vigorously and heated to 60°C. for 4 h, and then allowed to cool. The slurry was diluted with water,and acidified to pH=2 by addition of HCl. The resulting precipitate wascollected by filtration, washed with water, and dried under vacuum, toafford a 5-alkyl-4,6-dihydroxypyrimidine of sufficient purity forimmediate conversion to a 5-alkyl-4,6-dichloropyrimidine.

Method PP Preparation of 5-Alkoxy-4-hydroxypyrimidine

The method (p. 308) of Anderson et al. (Org. Proc. Res. Devel. 1997, 1,300-310) was employed to react a methyl alkoxyacetate, sodium methoxide,ethyl formate, and formamidine acetate to afford a5-alkoxy-4-hydroxypyrimidine of sufficient purity for immediateconversion to a 5-alkoxy-4-chloropyrimidine.

Method QQ Preparation of 5-Alkyl-4,6-dichloropyrimidine or5-Alkoxy-4-chloropyrimidine

To a 5-alkyl-4,6-dihydroxypyrimidine or a 5-alkoxy-4-hydroxypyrimidine(1.0 eq) were added phosphorus oxychloride (15.0 eq) andN,N-dimethylaniline (1.0 eq), and the mixture was heated to 100° C. for3 h, and then allowed to cool. The resulting solution was poured ontoice, and the mixture was extracted with dichloromethane. The organicextracts were treated with anhydrous magnesium sulfate, filtered, andevaporated to afford a 5-alkyl-4,6-dichloropyrimidine or a5-alkoxy-4-chloropyrimidine of sufficient purity for immediateconversion to a 5-alkyl-4-N-alkylamino-6-chloropyrimidine or a5-alkoxy-4-N-alkylaminopyrimidine.

Method RR Preparation of 5-Alkyl-4-N-alkylamino-6-chloropyrimidine or5-Alkoxy-4-N-alkylaminopyrimidine

To a solution of a 5-alkyl-4,6-dichloropyrimidine or a5-alkoxy-4-chloropyrimidine (1.0 eq) in ethanol were added an alkylamine (1.2 eq, typically L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester) and diisopropylethylamine (2.0 eq). The mixture wassealed in a pressure tube and heated to 120° C. for 48 h, at which pointTLC indicated consumption of the 5-alkyl-4,6-dichloropyrimidine or the5-alkoxy-4-chloropyrimidine. The mixture was evaporated, and the residuewas partitioned between ethyl acetate and pH=4.5 citrate buffer. Theorganic extracts were washed with saturated sodium chloride, treatedwith anhydrous magnesium sulfate, filtered, and evaporated. The residuewas purified by chromatography on silica gel using ethyl acetate andhexanes to afford a pure 5-alkyl-4-N-alkylamino-6-chloropyrimidine or5-alkoxy-4-N-alkylaminopyrimidine.

Method SS Preparation of 5-Alkyl-4-N-alkylaminopyrimidine (Procedure I)

A suspension of 5-alkyl-4-N-alkylamino-6-chloropyrimidine (1.0 eq), andan equal mass 10% palladium on carbon, and sodium bicarbonate (5.0 eq)in methanol was shaken under 55 psi hydrogen gas for 16 h, at whichpoint TLC indicated consumption of the5-alkyl-4-N-alkylamino-6-chloropyrimidine. The mixture was filteredthrough Celite and evaporated to give a residue, which was partitionedbetween ethyl acetate and 70% saturated sodium chloride. The organicextracts were treated with anhydrous magnesium sulfate, filtered, andevaporated. The residue was purified by chromatography on silica gelusing ethyl acetate and hexanes to afford a pure5-alkyl-4-N-alkylaminopyrimidine.

Method TT Preparation of 5-Alkyl-4-N-alkylaminopyrimidine (Procedure II)

A suspension of 5-alkyl-4-N-alkylamino-6-chloropyrimidine (1.0 eq),sodium acetate (10.0 eq), and zinc powder (20.0 eq) in a 9:1 mixture ofacetic acid and water was stirred vigorously at 40° C. for 72 h, atwhich point TLC indicated partial consumption of the5-alkyl-4-N-alkylamino-6-chloropyrimidine. The supernatant solution wasdecanted from remaining zinc and evaporated. The residue was partitionedbetween ethyl acetate and saturated sodium bicarbonate, and the organicextracts were treated with anhydrous magnesium sulfate, filtered, andevaporated. The residue was purified by chromatography on silica gelusing ethyl acetate and hexanes to afford a pure5-alkyl-4-N-alkylaminopyrimidine.

Method UU Preparation of N-Benzyloxycarbonyl-L-Tyrosine tert-Butyl Ester

To a 0° C. suspension of L-tyrosine tert-butyl ester (Bachem, 1.0 eq)and sodium bicarbonate (2.0 eq) in a 1:1 mixture of THF and water wasadded slowly with stirring benzyl chloroformate (1.1 eq). After theaddition, the mixture was stirred at 0° C. for 3 h and at roomtemperature for 24 h. The mixture was diluted with diethyl ether, andthe aqueous layer was separated. The organic extracts were washed withsaturated sodium chloride, treated with anhydrous magnesium sulfate,filtered, and evaporated to afford N-benzyloxycarbonyl-L-tyrosinetert-butyl ester of sufficient purity for immediate conversion of thetyrosine hydroxyl into a carbamate.

Method VV Preparation ofN-Benzyloxycarbonyl-L-4-(N,N-Dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

A mixture of N-benzyloxycarbonyl-L-tyrosine tert-butyl ester (1.0 eq),4-dimethylaminopyridine (1.0 eq), triethylamine (1.5 eq),dimethylcarbamylchloride (1.2 eq), and dichloromethane was heated to 37°C. for 16 h. The mixture was diluted with additional dichloromethane andwashed sequentially with 1.0 M potassium bisulfate, water, saturatedsodium bicarbonate, and saturated sodium chloride. The organic extractswere treated with anhydrous magnesium sulfate, filtered, and evaporatedto afford N-benzyloxycarbonyl-L-4-(N,N-dimethylcarbamyloxy)phenylaIaninetert-butyl ester as a white solid of sufficient purity for immediateconversion to L-4-(N,N-dimethylcarbamyloxy)phenylalanine tert-butylester.

Method WW Preparation of L-4-(N,N-Dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

A suspension ofN-benzyloxycarbonyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester and an equal mass of 10% palladium on carbon inmethanol was shaken under 55 psi hydrogen gas for 1 h, at which pointTLC indicated consumption of theN-benzyloxycarbonyl-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester. The mixture was filtered through Celite and evaporatedto afford L-4-(N,N-dimethylcarbamyloxy)-phenylalanine tert-butyl esterof sufficient purity for immediate use in reactions withchloropyrimidines.

Method XX Preparation ofN-Benzyloxycarbonyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalaninetert-Butyl Ester

To a stirred solution maintained at 0° C. ofN-benzyloxycarbonyl-L-tyrosine tert-butyl ester (1.0 eq) andtriethylamine (2.5 eq) in dichloromethane was added 4-nitrophenylchloroformate (1.0 eq). The mixture was stirred for 30 min at 0° C., andthen 1- methylpiperazine (1.5 eq) was added, and then the mixture wasstirred for 2 h while warming to room temperature. The mixture wasdiluted with ethyl acetate and washed five times with 10% potassiumcarbonate and once with saturated sodium chloride. The organic extractswere treated with anhydrous magnesium sulfate, filtered, and evaporatedto afford N-benzyloxycarbonyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalaninetert-butyl ester of sufficient purity for immediate conversion toL4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine tert-butyl ester.

Method YY Preparation ofL-4-(4-Methylpiperazin-1-ylcarbonyloxy)phenylalanine tert-Butyl Ester

A suspension ofN-benzyloxycarbonyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)-phenylalaninetert-butyl ester and an equal mass of 10% palladium on carbon inmethanol was shaken under 55 psi hydrogen gas for 1 h, at which pointTLC indicated consumption ofN-benzyloxycarbonyl-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalaninetert-butyl ester. The mixture was filtered through Celite and evaporatedto afford L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalaninetert-butyl ester of sufficient purity for immediate use in reactionswith chloropyrimidines.

Method ZZ tert-Butyl Ester Cleavage Procedure IV

The tert-butyl ester was dissolved in 96% formic acid and heated to 40°C. for 16 h, at which point TLC. indicated consumption of the tert-butylester. The mixture was evaporated under a stream of air to give aresidue, which was stored under high vacuum for 72 h to afford the purecarboxylic acid.

Method AAA Preparation of 2,4-Dichloro-5-nitropyrimidine

5-Nitrouracil was treated with phosphorus oxychloride andN,N-dimethylaniline, according to the procedure of Whittaker (J. Chem.Soc. 1951, 1565), to give 2,4-dichloro-5-nitropyrimidine as an orangeoil, which was used without distillation immediately in the next step.

Method BBB Preparation ofN-(2-Chloro-5-nitropyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

To a stirred solution of L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester (6.38 g, 20.69 mmol) and N,N-diisopropylethylamine(5.40 mL, 4.01 g, 31.03 mmol) in 70 mL CH₂Cl₂ at 0° C., was added asolution of 2,4-dichloro-5-nitropyrimidine (3.25 g, 20.69 mmol) in 70 mLof CH₂Cl₂, at such a rate the temperature did not exceed 10° C. Afterthe addition, the mixture was stirred at 0-10° C. for 15 minutes, atwhich point TLC indicated conversion of 2,4-dichloro-5-nitropyrimidine.To the mixture were added 100 mL 1 M KHSO₄ and 200 mL diethyl ether. Theorganic layer was separated, washed (H₂O, sat. NaHCO₃, and sat. NaCl),dried (MgSO₄), filtered, and evaporated to giveN-(2-chloro-5-nitropyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester (9.52 g, 20.45 mmol, 99%) as an orange oil, which wasused immediately in the next step.

Method CCC Preparation ofN-(5-Aminopyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

A mixture ofN-(2-chloro-5-nitropyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester (9.52 g, 20.45 mmol), Degussa-type 20% palladium oncarbon (9.52 g), NaHCO (8.59 g, +02.2 mmol) and 165 mL MeOH was shakenunder 55 psi H₂ for 16 h, at which point TLC indicated conversion ofN-(2-chloro-5-nitropyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester into a single product. The mixture was filtered throughCelite, and the filtrate was evaporated to give a residue, which wasdissolved by addition of 150 mL EtOAc and 75 mL H₂O. The organic layerwas separated, washed (sat. NaCl), dried (MgSO₄), filtered, andevaporated to giveN-(5-aminopyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester (7.14 g, 17.79 mmol, 87%) as an orange solid, which wasused immediately in the next step.

Method DDD Preparation ofN-(5-(N-4-Toluenesulfonylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)pbenylalaninetert-Butyl Ester

To a stirred solution ofN-(5-aminopyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester (1.00 g, 2.49 mmol) in 10 mL anhydrous pyridine at 0°C., was added in portions 4-toluenesulfonylchloride (0.474 g, 2.49mmol). After the addition, the resulting red solution was stirred at 0°C. for 3 h, at which point TLC indicated nearly complete conversion ofN-(5-aminopyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester. To the mixture was added 3-dimethylaminopropylamine(0.325 mL, 0.264 g, 2.49 mmol), and the mixture was stirred for 30 minwhile warming to room temperature. The mixture was poured into 100 mL 1M KHSO₄, and extracted with 150 mL EtOAc. The organic layer was washed(2×1 M KHSO₄, H₂O, sat. NaHCO₃, sat. NaCl), dried (MgSO₄), filtered, andevaporated to give a brown residue, which was purified by flashchromatography using EtOAc/hexanes on silica gel, to giveN-(5-(N-4-toluenesulfonylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester (1.01 g, 1.81 mmol, 73%) as a clear oil.

Method EEE Preparation ofN-(5-(N-Methyl-N-4-toluenesulfonylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine tert-Butyl Ester

To a stirred two-phase mixture of 45 mL 1 M NaOH and 25 mL diethyl etherat 0° C., was added in portions 1-methyl-3-nitro-1-nitrosoguanidine(1.33 g, 9.05 mmol). After stirring for 25 min, at which point evolutionof N₂ had subsided, the bright yellow solution-of diazomethane indiethyl ether was transferred by pipette to a stirred solution ofN-(5-(N-4-toluenesulfonylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester (1.01 g, 1.81 mmol) in 15 mL diethyl ether and 15 mLCH₂Cl₂ at 0° C. After stirring for 15 min, at which point TLC indicatedcomplete conversion ofN-(5-(N-4-toluenesulfonylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester, excess AcOH was added to destroy unreacteddiazomethane. The mixture was diluted with 100 mL diethyl ether, washed(2× sat. NaHCO₃, sat. NaCl), dried (MgSO₄), filtered and evaporated togive a yellow residue, which was purified by flash chromatography usingEtOAc/hexanes on silica gel, to giveN-(5-(N-methyl-N-4-toluenesulfonylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester (0.846 g, 1.48 mmol, 82%) as a clear oil.

Method FFF

Preparation of Diethyl 2-(N,N-Dialkylamino)malonate

The appropriate amine (1.0 eq) was added to a 0° C. solution of diethylbromomalonate (1.0 eq) and N,N-diisopropylethyl amine (1 .1 eq) inethanol. The mixture was stirred and allowed to warm room temperature.After 16 hours, the reaction mixture was concentrated and the residuewas suspended in ethyl acetate and sat. NaHCO₃. The organic portion waswashed with sat NaHCO₃, brine, dried (MgSO₄) filtered and concentratedto yield the diethyl 2-(N,N-dialkylamino)malonate, of sufficient purityfor immediate conversion to a5-(N,N-dialkylamino)-4,6-dihydroxypyrimidine.

Method GGG Preparation of 5-(N,N-Dialkylamino)-4,6-dihydroxypyrimidine

A suspension of a diethyl 2-(N,N-dialkylamino)malonate (1.0 eq),formamidine acetate (1.10 eq.) and 25% sodium methoxide in methanol (3.3eq) was heated to 65° C. for 3.5 hours. The reaction mixture was cooledand diluted with water. The mixture was acidified to pH=4.5 by additionof dilute HCl. The resulting precipitate was collected by filtration,washed with water, and dried under vacuum to afford a5-(N,N-dialkylamino)-4,6-dihydroxypyrimidine of sufficient purity forimmediate conversion to a 5-(N,N-dialkylamino)-4,6-dichloropyrimidine.Alternatively, the acidified solution was evaporated to give a solidresidue, which was extracted with boiling ethanol. The ethanol extractswere filtered and concentrated to give a residue, which wasrecrystallized from isopropyl alcohol to afford a5-(N,N-dialkylamino)-4,6-dihydroxypyrimidine of sufficient purity forimmediate conversion to a 5-(N,N-dialkylamino)-4,6-dichloropyrimidine.

Method HHH Preparation of 5-(N.N-Dialkylamino)-4,6-dichloropyrimidine

A 5-(N,N-dialkylamino)-4,6-dihydroxypyrimidine (1.0 eq) was suspended inPOC13 (15.0 eq), and the mixture was heated to reflux for 16 hours. Thenthe mixture was cooled and carefully poured into a suspension of ethylether and aqueous K₂CO₃. The organic portion was washed with brine,dried (MgSO₄), filtered and concentrated to yield a 5-(N,N-dialkylamino)-4,6-dichloro-pyrimidine of sufficient purity for immediatereaction with alkylamines.

Method III Preparation of4-(N-Alkylamino)-5-(N,N-dialkylamino)-6-chloropvrimidine

A 5-(N,N-dialkylamino)-4,6-dichloropyrimidine (1.0 eq),L-4-(N,N-dimethylcarbamyloxy)phenylalanine tert-butyl ester (1.5 eq) andN,N-diisopropyl ethylamine (1.5 eq) were dissolved in ethanol and heatedto 120° C. in a sealed tube for 72 h. The cooled reaction mixture wasconcentrated, and the residue dissolved in ethyl acetate. The ethylacetate solution was washed with 0.2 N citric acid, water, saturatedNaHCO₃, brine, dried (MgSO₄), filtered and concentrated. The residue waspurified by silica gel chromatography using ethyl acetate/hexanes toafford the 4-(N-alkylamino)-5-(N,N-dialkylamino)-6-chloropyrimidine.

Method JJJ Preparation of4-(N-Alkylamino)-5-(N,N-dialkylamino)pyrimidine

A 4-(N-Alkylamino)-5-(N,N-dialkylamino)-6-chloropyrimidine (1.0 eq), anequal mass of 10% palladium on carbon, and NaHCO₃ (5.0 eq) weresuspended in methanol. The reaction mixture was hydrogenated at 45 psihydrogen for 16 hours and then filtered through a pad of Celite. Thefiltrate was concentrated, and the residue was dissolved in ethylacetate. The ethyl acetate solution was washed with water, brine, dried(MgSO₄), filtered and concentrated to yield an oil. The oil was purifiedby column chromatorgraphy on silica gel using ethyl actate and hexanesto afford a pure 4-(N-alkylamino)-5-(N,N- dialkylamino)pyrimidine.

Method KKK Suzuki Coupling Procedure V

To an ethyleneglycol dimethyl ether solution oftetrakis(triphenylphosphine) palladium (0.04 eq) was addedN-(5-bromo-2-chloro-pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester (1.5 eq.). After stirring for approximately ten minuteso-tolylboronic acid (1.5 eq) and 2M Na₂CO₃ (2.0 eq) were added, and thereaction flask was evacuated and flushed with nitrogen gas. The reactionwas heated tp reflux for four hours. The reaction mixture was thencooled and diluted with water and methylene chloride. The organic phasewas separataed and washed with brine, dried (MgSO₄), filtered andconcentrated. The residue was purified by silica gel chromatographyusing ethyl acetate/hexanes to afford the desired product.

Method LLL Preparation of L-Phenylalanine Isopropyl Ester Hydrochlorideor L-Tyrosine Isopropyl Ester Hydrochloride

Excess HCl gas was added with stirring to a suspension ofL-phenylalanine or L-tyrosine in excess isopropanol. The mixture washeated to reflux for 16 h, and then the volatiles were evaporated undervacuum to give L-phenylalanine isopropyl ester hydrochloride orL-tyrosine isopropyl ester hydrochloride of sufficient purity forimmediate use.

Method MMM Bromopyrimidine Debromination Procedure

The bromopyrimidine was dissolved in isopropyl alcohol to which wasadded 10% palladium on carbon. The reaction was hydrogenated at 45 psihydrogen. Filtration and concentration of the filtrate yielded thedesired dehalogenated pyrimidine.

Method NNN Preparation of 2-Isopropropoxypyrimidine

A 2-chloropyrimidine was dissolved in isopropyl alcohol to which wasadded diisopropylamine. The reaction was heated in a sealed tube for tendays at 130° C. The cooled reaction mixture was concentrated, and theproduct purified via silica gel column chromatography to yield the2-isopropoxypyrimidine.

Method OOO Heck Reaction Procedure III

To a dioxane/triethylamine (1:1 by volume) solution of theN-(5-iodopyridin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanineisopropyl ester (1.0 eq), triphenylphosphine (0.05 eq), copper (I)iodide (0.2 eq) was added phenylacetylene (4.0 eq). After flushing thesolution for ten minutes with nitrogen gas,dichlorobis(triphenylphosphine)palladium (0.10 eq) was added, and theresulting reaction mixture heated to 50° C. for 16 hours. The reactionmixture was then diluted with ethyl acetate and water, and the organicportion was washed with 0.2 N citric acid, water, saturated NaHCO₃,brine, dried (MgSO₄), filtered and concentrated. The residue waschromatographed on a silica gel column using ethyl acetate/hexanes toafford the desired product.

Method PPP Preparation ofN-5-(Phenyl)pyrimidin-4-yl]-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert- Butyl Ester

N-[5-iodopyrimidin-4-yl]-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester (123 mg, 0.2 mmol) was diluted in dry DMF (5 mL) undernitrogen with KOAc (3.0 eq, 73 mg), bis(pinacolato)diboron (1.1 eq, 63mg), and a catalytic amount of [1,1′-bis(diphenylphosphino)-ferrocene]dichloropalladium (II) complex withdichloromethane (1:1). The reaction was heated for 2 hours at 100° C. Tothis was added, K3PO4 (2.0 eq, 105 mg), iodobenzene (2.0 eq, 0.056 mL)and an additional catalytic amount of[1,1′-bis(diphenylphosphino)-ferrocene]dichloropalladium (II) complexwith dichloromethane (1:1). The reaction mixture was stirred overnightat 100° C. EtOAc was added and the organic layer washed with brine,dried over MgSO4. Upon filtration, and evaporation of the solvent underreduced pressure, the crude material was eluted on column chromatography(silica gel) with EtOAc/hexanes 1:1. The desired material was isolatedin good yields.

Method QQQ Preparation of 2-Amino-3-Chloropyrazine

A mixture of 2,3-dichloropyrazine (Lancaster) and ammonium hydroxide washeated in a sealed tube at 100° C. for 24 h resulting in a whiteprecipitate. The precipitate was collected by filtration and dried undervacuum to afford 2-amino-3-chloropyrazine of sufficient purity forimmediate conversion to 2-chloro-3-nitropyrazine.

Method RRR Preparation of 2-Chloro-3-Nitropyrazine

The method (p. 1638) of Hartman et al. (J. Med. Chem. 1984, 27(12),1634-1639) was employed to convert 2-amino-3-chloropyrazine into2-chloro-3-nitropyrazine of sufficient purity for immediate use.

Method SSS Preparation of 4-Alkylamino-2-dialkylamino-5-nitropyrimidine

A solution of 1.0 eq 4-alkylamino-2-chloro-5-nitropyrimidine and 5.0 eqdialkylamine in THF was allowed to stand for 16 h. The mixture wasdiluted with ethyl acetate and then washed with pH =4.5 citrate bufferand saturated sodium chloride. The organic extracts were treated withanhydrous magnesium sulfate, filtered, and evaporated to give a residue,which was purified by chromatography on silica gel using ethyl acetateand hexanes.

Method TTT Preparation of L-4-(2,6-Dimethoxyphenyl)phenylalanine MethylEster

To a stirred solution (DMF, 66 mL) of N-Boc-L-(p-iodo)phenylalaninemethyl ester (13.2 g, 32.7 mmol) prepared according to the procedure ofSchwabacher et al., J. Org. Chem. 1994, 59, 4206-4210) was addedPd(PPh₃)₄ (0.03 eq, 1.13 g, 1 mmol). The solution was stirred for 10 minand then 2,6-dimethoxyboronic acid (1.2 eq, 7.1 g, 39 mmol) and K₃PO₄(1.5 eq, 10.4 g, 49 mmol) were added. The reaction flask was evacuatedand flushed with nitrogen. This process was repeated twice and thereaction mixture was then heated to 100° C. under a stream of nitrogenfor about 3.5 h at which time TLC showed the reaction to be complete(4.5:1 hexanes:EtOAc, R_(f)=0.2, UV active). The reaction mixture wascooled and partitioned between water and ethyl acetate (200 mL each).The organic portion was washed with 0.2N citric acid (3×100 mL), brine(1×100 mL), dried (MgSO₄), filtered and stripped to a thick reddish oil,about 13 g. The resulting product was chromatographed on silica geleluting with 4.5:1 hexanes/EtOAc, R_(f)=0.2. The combined fractions werestripped and treated with methanol saturated with HCl to yield the titleintermediate as the hydrochloride salt.

Example 382 Synthesis ofN-(2-Chloro-5-nitropyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Step A—Preparation of 2,4-Dichloro-5-nitropyrimidine 5-Nitrouracil(Aldrich Chemical Company) was treated with phosphorous oxychloride andN,N-dimethylaniline according to the procedure described in Whittaker,J. Chem. Soc. 1951, 1565, to give 2,4-dichloro-5-nitropyrimidine as anorange oil which was used without distillation immediately in the nextstep.

Step B—Preparation ofN-(2-Chloro-5-nitropvrimidin-4-yl)-L-4-(N,N-dimethylcarbamvloxy)phenylalaninetert-Butyl Ester

To a stirred solution of L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester (6.38 g, 2069 mol) and N,N-diisopropylethylamine (5.40mL, 4.01 g, 31.03 mol.) in 70 mL CH₂Cl₂ at 0° C., was added a solutionof 2,4-dichloro-5-nitropyrimidine (3.25 g, 20.69 mol.) in 70 mL CH₂Cl₂at such a rate that the temperature did not exceed 10° C. After theaddition, the mixture was stirred at 0- 10° C. for 15 minutes, at whichpoint TLC indicated conversion of the starting materials. To the mixturewere added 100 mL 1 M KHSO4 and 200 mL diethyl ether. The organic layerwas separated, washed (H₂O, sat. NaHCO₃, and sat. NaCl), dried (MgSO₄),filtered, and evaporated to give the title compound (9.52 g, 2045 mol.,99%) as an orange oil.

Step C—Preparation ofN-(2-Chloro-5-nitropvrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared by hydrolysis of the product from Step Busing the procedure of Example 386.

Example 383 Synthesis ofN-[5-(N-4-Toluenesulfonylamino)pyrimidin-4-yll-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

Step A—Preparation ofN-(5-Aminopyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

A mixture ofN-(2-chloro-5-nitropyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-butyl ester (9.52 g, 20.45 mol), Degussa-type 20% palladium oncarbon (9.52 g), NaHCO₃ (8.59 g, 102.2 mol), and 165 mL MeOH was shakenunder 55 psi for 16 h, at which point TLC indicated conversion of thestarting material into a single product. The mixture was filteredthrough Celite, and the filtrate was evaporated to give a residue, whichwas dissolved by addition of 150 mL EtOAc and 75 mL H₂O. The organiclayer was separated, washed (sat. NaCl), dried (MgSO₄), filtered, andevaporated to give the title intermediate (7.14 g, 17.79 mol, 87%) as anorange solid, which was used immediately in the next step.

Step B—Preparation ofN-F5-(N-4-Toluenesulfonyl-amino)pyrimidin-4-yl]-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

To a stirred solution of the product from Step A (100 g, 2.49 mol) in 10mL anhydrous pyridine at 0° C., was added in portions 4-toluenesulfonylchloride (0.474 g, 2.49 mol). After the addition, the resulting redsolution was stirred at 0° C. for 3 h, at which point TLC indicatednearly complete conversion of the starting material. To the mixture wasadded 3-dimethylaminopropylamine (0.325 mL, 0.264 g, 2.49 mol), and themixture was stirred for 30 min while warming to room temperature. Themixture was poured into 100 mL 1 M KHSO4, and extracted with 150 mLEtOAc. The organic layer was washed (2×1 M KHSO4, H₂O, sat. NaHCO₃, sat.NaCl), dried (MgSO₄), filtered, and evaporated to give a brown residue,which was purified by flash chromatography using EtOAc/hexanes on silicagel, to give the title compound (1.01 g, 1.81 mol., 73%) as a clear oil.

Example 384 Synthesis ofN-15-(N-4-Toluenesulfonylamino)pyrimidin-4-yl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The title compound was prepared by hydrolysis of-N-[5-(N-4-toluenesulfonylamino)pyrimidin-4-yl]-L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert- butyl ester using the procedure of Example 386.

Example 385 Synthesis of N-15-(N-Methyl-N-4-toluenesulfonylamino)pyrimidin-4-yl]-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

To a stirred two-phase mixture of 45 mL 1 M NaOH and 25 mL.diethyl etherat 0° C., was added in portions 1-methyl-3-nitro-1-nitrosoguanidine(1.33 g, 9.05 mol). After stirring for 25 min, at which point evolutionof N₂ had subsided, the bright yellow solution of diazomethane indiethyl ether was transferred by pipette to a stirred solution of theproduct of Example 383 (1.01 g, 1.81 mol) in 15 mL diethyl ether and 15mL CH₂Cl₂ at 0° C. After stirring for 15 min, at which point TLCindicated complete conversion of the starting material, excess AcOH wasadded to destroy unreacted diazomethane. The mixture was diluted with100 mL diethyl ether, washed (2× sat. NaHCO₃, sat. NaCl), dried (MgSO₄),filtered and evaporated to give a yellow residue, which was purified byflash chromatography using EtOAc/hexanes on silica gel, to give thetitle compound (0.846 g, 1.48 mol, 82%) as a clear oil.

Example 386 Synthesis of N-[5-(N-Methyl-N-4-toluenesulfonylamino)pyrimidin-4-yl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The product of Example 385 (0.400 g, 0.700 mol) was dissolved in 8 mL96% formic acid, and the mixture was heated to 40° C. for 16 h, at whichpoint TLC indicated conversion of the starting material. Most of theformic acid was evaporated under a stream of N₂, and then the residuewas placed under high vacuum for 48 h to give the title compound (0.382g, 0.700 mol, 100%) as a clear oil.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.33 (bs, 1H), 8.07 (bs, 1H), 7.64 (d, J=8.1 Hz, 2H),7.42 (d, J=8.1 Hz, 2H), 7.36 (bs, 1H), 7.29 (bs, 2H), 6.99 (d, J=7.5 Hz,2H), 5.07-4.96 (m, 1H), 3.42-3.31 (m, 1H), 3.25-3.15 (m, 1H), 3.08 (s,3H), 3.05 (bs, 3H), 2.96 (s, 3H), 2.44 (s, 3H).

¹³C NMR (CD₃OD): δ=174.7, 174.6, 164.6, 157.8, 156.8, 152.9, 152, 146,35.4, 135.1, 131.7, 131.3, 129.4, 123.2, 122.9, 55.8, 38.2, 37.1, 36.8,36.7, 21.5.

Using the appropriate starting materials and reagents, the followingadditional compounds were prepared:

-   N-[5-(N,N-Di-4-toluenesulfonylamino)pyrimidin-4-yl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    (Example 387);-   N-[5-[N-(1-A-Methylpyrazol-4-ylsulfonyl)-N-methylamino]pyrimidin-4-yl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    (Example 388);-   N-[5-(N-Methyl-N-4-toluenesulfonylamino)pyrimidin-4-yl]-L-4-(N,N-    dimethylcarbamyloxy)phenylalanine isopropyl ester (Example 389);-   N-[5-(N-Methyl-N-3 -pyridylsulfonylamino)pyrimidin-4-yl]-L-4-(N,N-    dimethylcarbamyloxy)phenylalanine tert-butyl ester (Example 390).

Example 391 Synthesis ofN-(5-(N-Methyl-N-(1-butylpyrazol4-yl)sulfonylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 5-Nitrouracil (Aldrich) was converted via Method AAAinto 2,4-dichloro-5-nitropyrimidine.L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butyl ester and2,4-dichloro-5-nitropyrimidine were coupled via Method BBB, and theproduct of this coupling was sequentially converted via Methods CCC, DDD(using 1 -butyl-4-chlorosulfonylpyrazole), EEE and ZZ to give the titlecompound.

Physical data were as follows:

¹H NMR ( CD₃OD ): δ=8.35 (s, 1H), 8.14 (s, 1H), 7.76 (s, 1H), 7.61 (bs,1H), 7.23 (bs, 2H), 6.98 (d, 2H), 5.01-4.94 (m, 1H), 4.19 (t, 2H),3.40-3.28 ( m, 1H ), 3.26-3.14 (m,1H), 3.09 (s, 3H), 3.06 (bs, 3H), 2.96(s, 3H), 1.84 (pent., 2H), 1.29 (sext., 2H), 0.945 (t, 3H).

Example 392 Synthesis ofN-(5-(2,4-Dimethoxypyrimidin-5-yl)pyrimidin4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 4(3H)-pyrimidinone (Aldrich) was sequentiallyconverted via Methods N and O into 4-chloro-5-iodopyrimidine.L-4-(N,N-dimethylcarbamyloxy)phenylalanine tert-butyl ester and4-chloro-5-iodopyrimidine were coupled via Method P, and the coupledproduct was reacted with 2,4-dimethoxypyrimidin-5-yl boronic acid(Frontier Scientific, Inc.) via Method S. The product of this couplingwas converted via Method KK to give the title compound.

Example 393 Synthesis ofN-(5-(2,6-Difluorophenyl)pyridin4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 4(3H)-Pyrimidinone (Aldrich) was sequentiallyconverted via Methods N and O into 4-chloro-5-iodopyrimidine.L-4-(N,N-Dimethylcarbamyloxy)phenylalanine tert-butyl ester and4-chloro-5-iodopyrimidine were coupled via Method P, and the coupledproduct was reacted with 2,6-difluorophenyl boronic acid (LancasterSynthesis) via Method R. The product of this coupling was converted viaMethod HH to give the title compound.

Example 394 Synthesis ofN-(5-(2-Hydroxymethylphenyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 4(3H)-Pyrimidinone (Aldrich) was sequentiallyconverted via Methods N and O into 4-chloro-5-iodopyrimidine.L-4-(N,N-dimethylcarbamyloxy)phenylalanine tert-butyl ester and4-chloro-5-iodopyrimidine were coupled via Method P, and the coupledproduct was reacted with 2-(hydroxymethyl)phenyl boronic acid (LancasterSynthesis) via Method Q. The product of this coupling was converted viaMethod HH to give the title compound.

Example 395 Synthesis ofN-(2-(N-Cyclohexylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted with cyclohexylamine(Aldrich) via Method CC to give a product that was coupled with o-tolylboronic acid (Aldrich) via Method DD. The product of this coupling wasconverted via Method ZZ to give the title compound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=9.68 (s, 1H), 7.3-6.8 (m, 9H), 6.35 (m, 1H), 4.73(m,1H), 3.81 (bs, 1H), 3.6-3.0 (m, 2H), 3.09 (s, 3H), 3.0 (s, 3H), 2.18 (s,1.5H), 1.94 (s, 1.5H), 2.1-1.1 (m, 10H).

¹³C NMR (CDCl₃): δ=176.11, 175.94, 160.05, 159.79, 154.76,153.58,150.05, 150.01, 139.26, 137.84, 137.63, 134.29, 134.15, 130.66, 130.36,130.11, 129.14, 126.70, 126.41, 121.25, 109.57, 109.39, 56.84, 56.35,50.15, 36.55, 36.32, 32.34, 31.99, 25.41, 24.86, 19.48, 19.27.

Example 396 Synthesis ofN-(2-(N-Methyl-N-(1-methylpiperidin-4-yl)amino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted with 1-methyl-4-(N-methylamino)piperidine (Aldrich) via Method CC to give aproduct that was coupled with o-tolyl boronic acid (Aldrich) via MethodDD. The product of this coupling was converted via Method ZZ to give thetitle compound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=8.82 (s, 2H), 8.43 (s, 1H), 7.62 (s, 1H), 7.30-6.90(m, 8H), 5.42 (br, 1H), 4.66 (br, 2H), 3.60-2.8 (m, 15H), 2.66 (bs, 3H),2.32 (br, 2H), 2.18 (s, 1.5H), 1.82 (brs, 3.5H).

Example 397 Synthesis ofN-(2-(N-Ethyl-N-isopropylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted withN-ethyl-N-isopropylamine (Aldrich) via Method CC to give a product thatwas coupled with o-tolyl boronic acid (Aldrich) via Method DD. Theproduct of this coupling was converted via Method ZZ to give the titlecompound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=8.0-6.5 (br, 1H), 7.66 (s, 0.5H), 7.62 (s, 0.5H),7.3-6.8 (m, 8H), 6.2 (m, 1H), 4.86 (br, 1H), 4.70 (m, 1H), 3.70-3.08 (m,4H), 3.09 (s, 3H), 3.0 (s, 3H), 2.14 (bs, 1.5H), 1.92 (bs, 1.5H),1.4-0.9 (br, 9H).

¹³C NMR (CDCl₃): δ=174.38, 174.19, 159.44, 159.16, 155.24, 154.68,152.39, 150.02, 141.63, 137.77, 137.56, 134.30, 134.09, 130.79, 130.66,130.54, 130.46, 130.41, 130.33, 130.08, 129.07, 126.54, 126.45, 126.38,121.21, 121.16, 110.27, 110.01, 56.77, 56.36, 47.59, 36.80, 36.55,36.32, 20.27, 20.18, 19.57, 19.38, 14.51.

Example 398 Synthesis ofN-(5-(2,4-6-Trimethylphenyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 4(3H)-Pyrimidinone (Aldrich) was sequentiallyconverted via Methods N and O into 4-chloro-5-iodopyrimidine.L-4-(N,N-Dimethylcarbamyloxy)phenylalanine tert-butyl ester and4-chloro-5-iodopyrimidine were coupled via Method P, and the coupledproduct was reacted with 2,4,6-trimethylphenyl boronic acid (FrontierScientific, Inc) via Method R. The product of this coupling wasconverted via Method HH to give the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.68 (d, 1H), 7.95 (d, 1H), 7.10 (d, 2H), 7.09-6.95(m,2H), 6.94-6.91 (m, 2H), 5.32-5.27 (m, 1H), 3.42-3.36 (m, 1H), 3.15-3.09(m, 4H), 2.97 (s, 3H), 2.33 (s, 3H), 2.04 (s, 3H), 1.84 (s, 3H).

¹³CNMR(CD₃OD): δ=172.9, 163.5, 161.5, 161.0, 156.7, 152.0, 151.9, 142.6,141.5, 138.9, 138.6, 135.3, 131.2, 130.4, 130.3, 126.5, 123.0, 120.3,56.4, 36.7,36.6, 36.5, 21.2, 19.9, 19.7.

Example 399 Synthesis ofN-(5-Isopropylpyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. Diethyl 2-isopropylmalonate (Aldrich) was sequentiallyconverted via Methods OO and QQ into 4,6-dichloro-5-isopropylpyrimidine.L-4-(N,N-Dimethylcarbamyloxy)phenylalanine tert-butyl ester and4,6-dichloro-5-isopropylpyrimidine were coupled via Method RR, and theproduct of this coupling was sequentially converted via Methods SS andZZ to give the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.44 (bs, 1H), 7.94 (bs, 1H), 7.22 (d, 2H), 6.94(d,2H), 5.12 (dd, 1H), 3.46 (dd, 1H), 3.19 (dd, 1H), 3.07 (s, 3H), 2.95 (s,3H), 3.00-2.88 (m, 1H), 1.25 (d, 3H), 1.13 (d, 3H).

¹³C NMR (CD₃OD): δ=175.60, 165.74,163.78, 156.91, 152.38, 151.85,141.88, 136.30, 131.43, 126.17, 122.87, 57.84, 37.48, 36.81, 36.64,26.63, 21.09, 20.94.

Example 400 Synthesis ofN-(2-(N-Methyl-N-butylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dirnethylcarbamyloxy)-phenylalaninetert-butyl ester and 5-bromo-2,4-dichloropyrimidine (Aldrich) werecoupled via Method BB. The product of this reaction was reacted withN-methyl-N-butylamine (Aldrich) via Method CC to give a product that wascoupled with o-tolyl boronic acid (Aldrich) via Method DD. The productof this coupling was converted via Method ZZ to give the title compound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=12.5-11.4 (br, 1H), 7.6 (s, 0.5H), 7.58 (s, 0.5H),7.3-6.8 (m, 8H), 6.3 (m, 1H), 4.7 (m, 1H), 3.7-2.9 (m, 4H), 3.08 (s,3H), 3.01 (s, 6H), 2.13 (s, 1.5H), 1.91 (s, 1.5H), 1.57 (bs, 2H), 1.33(m, 2H), 0.96 (t, 3H).

¹³C NMR (CDCl₃): δ=174.21, 174.06, 159.37, 159.22, 154.69, 153.52,169.99, 141.87, 137.77, 137.54, 134.43, 130.78, 130.59, 130.10, 128.98,126.51, 126.32, 121.17, 121.11, 110.20, 109.96, 56.82, 56.43, 50.03,36.54, 36.32, 35.91, 29.27, 19.89, 19.52, 19.35, 13.84.

Example 401 Synthesis ofN-(2-(N-Ethyl-N-propylamino)-5-(2-tolyl)pyrimidin4-yi)-L4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted withN-ethyl-N-propylamine (Aldrich) via Method CC to give a product that wascoupled with o-tolyl boronic acid (Aldrich) via Method DD. The productof this coupling was converted via Method ZZ to give the title compound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=11.0-9.5 (br, 1H), 7.66 (s, 0.5H), 7.64 (s, 0.5H),7.4-6.8 (m, 8H), 6.28 (m, 1H), 4.65 (m, 1H), 3.70-2.80 (m, 6H), 3.09 (s,3H), 3.01 (s, 3H), 3.01 (s, 3H), 2.2 (s, 1.5H), 1.85 (s, 1.5H), 1.58(bs, 2H), 1.05 (bs, 3H), 0.85 (bs, 3H).

¹³C NMR (CDCl₃): δ=174.26, 174.11, 159.36, 159.11, 154.70, 153.07,149.96, 142.43, 137.80, 137.56, 134.54, 134.37, 130.84, 130.74, 130.57,130.14, 128.86, 126.47, 126.29, 121.10, 121.06, 110.01, 109.71, 56.86,56.49, 49.62, 63.20, 36.55, 36.32, 20.87, 19.61, 19.41, 12.63, 11.03.

Example 402 Synthesis ofN-(2-(N,N-Diethylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted withN,N-diethylamine (Aldrich) via Method CC to give a product that wascoupled with o-tolyl boronic acid (Aldrich) via Method DD. The productof this coupling was converted via Method ZZ to give the title compound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=12.2 (br, 1H), 7.63 (s, 0.5H), 7.60 (s, 0.5H),7.40-6.80 (m, 8H), 6.28 (m, 1H), 4.70 (m, 1H), 3.80-2.90 (m, 6H), 3.06(s, 3H), 2.98(s, 3H), 2.13 (s, 1.5H), 1.92 (s, 1.5H), 0.90 (s, 6H).

¹³C NMR (CDCl₃): δ=174.34, 174.15, 159.4, 159.1, 154.70, 152.66, 169.97,142.06, 137.76, 137.55, 134.44, 134.27, 130.81, 130.57, 130.10, 128.95,126.48, 126.32, 121.14, 121.08, 110.08, 109.80, 56.78, 56.37, 42.77,36.53, 36.31, 19.57, 19.38, 12.77.

Example 403 Synthesis ofN-(2-(N-Methyl-N-ethylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted withN-methyl-N-ethylamine (Aldrich) via Method CC to give a product that wascoupled with o-tolyl boronic acid (Aldrich) via Method DD. The productof this coupling was converted via Method ZZ to give the title compound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=12.5 (br, 2H), 8.23 (s, 1H), 7.50 (s, 0.5H), 7.44(s,0.5H), 7.30 6.80 (m, .8H),-6.10 (m, 1H),-4.75 (m, 1H), 3.58-(bs, 2H),3.30 (m, -H), 3.00-(m, 1H), 3.08 (s, 3H), 3.00 (s, 3H), 2.93(s, 3H),2.08 (s, 1.5H), 1.92 (s, 1.5H), 1.50 (s, 3H).

¹³C NMR (CDCl₃): δ=174.63, 174.34, 165.72, 159.96, 159.72, 154.88,152.62, 150.49, 150.45, 140.64, 137.90, 137.81, 133.83, 133.65, 131.03,130.95, 130.85, 130.63, 130.10, 130.04, 129.76, 129.62, 126.88, 126.72,121.70, 121.61, 110.69, 110.46, 56.65, 56.11, 45.16, 36.57, 36.35,35.17, 19.38, 19.17, 11.96.

Example 404 Synthesis of N-(5-Benzyloxypyrirnidin-4-yl)-L-phenylalanine

Methyl 2-benzyloxyacetate (Aldrich) was sequentially converted viaMethods PP and QQ into 4-chloro-5-benzyloxypyrimidine. L-4-phenylalaninetert-butyl ester (Bachem) and 4-chloro-5-benzyloxypyrimidine werecoupled via Method PP, and the product of this coupling was convertedvia Method ZZ to give the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.54 (s, formate), 8.03 (s, 1H), 7.67 (s, 1H),7.37-7.31(m, 5H), 7.17-7.12 (m, 5H), 5.11 (s, 2H), 4.78-4.75 (m, 1H),3.35-3.11 (m, 2H).

¹³C NMR (CD₃OD): δ=159.07, 143.16, 132.35, 130.64, 124.52, 123.94,123.83, 123.59, 123.11, 122.00, 99.47, 66.28, 50.32, 32.05.

Example 405 Synthesis ofN-(5-Benzyloxypyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. Methyl 2-benzyloxyacetate (Aldrich) was sequentiallyconverted via Methods PP and QQ into 4-chloro-5-benzyloxypyrimidine.L-4-(N,N-Dimethylcarbamyloxy)phenylalanine tert-butyl ester and4-chloro-5-benzyloxypyrimidine were coupled via Method RR, and theproduct of this coupling was converted via Method ZZ to give the titlecompound.

Example 406 Synthesis ofN-(5-(N-Methyl-N-4-toluenesulfonylamino)pyrimidin-4-yl)-L-phenylalanine

5-Nitrouracil (Aldrich) was converted via Method AAA into2,4-dichloro-5-nitropyrimidine. L-4-Phenylalanine tert-butyl ester(Bachem) and 2,4-dichloro-5-nitropyrimidine were coupled via Method BBB,and the product of this coupling was sequentially converted via MethodsCCC, DDD, EEE and ZZ to give the title compound.

Example 407 Synthesis ofN-(5-(N-Methyl-N-3-pyridinesulfonylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 5-Nitrouracil (Aldrich) was converted via Method AAAinto 2,4-dichloro-5-nitropyrimidine.L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butyl ester and2,4-dichloro-5-nitropyrimidine were coupled via Method BBB, and theproduct of this coupling was sequentially converted via Methods CCC, DDD(using 3-chlorosulfonylpyridine), EEE and ZZ to give the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.90 (d, 1H), 8.85 (d, 1H), 8.36 (s, 1H), 8.15 (d,1H), 7.64 (dd, 1H), 7.53 (bs, 1H), 7.27 (bs, 2H), 6.99 (d, 2H),5.04-4.87 (m, 1H), 3.40-3.28 (m, 1H), 3.26-3.16 (m, 1H), 3.13 (bs, 3H),3.09 (s, 3H), 2.97 (s, 3H).

Example 408 Synthesis ofN-(5-Phenylpyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 4(3H)-Pyrimidinone (Aldrich) was sequentiallyconverted via Methods N and O to 4-chloro-5-iodopyrimidine.L-4-(N,N-Dimethylcarbamyloxy)phenylalanine tert-butyl ester and4-chloro-5-iodopyrimidine were coupled via Method P, and the coupledproduct was reacted with phenyl boronic acid (Aldrich) via Method S. Theproduct of this coupling was converted via Method HH to give the titlecompound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.62 (s, 1H), 8.04 (s, 1H), 7.53-7.51 (m, 3H),7.30-7.27 (m, 2H), 7.17-7.15 (m, 2H), 7.00-6.97 (m, 2H), 5.27-5.22 (m,1H), 3.45-3.39 (m, 1H), 3.16-3.08 (m, 4H), 2.96 (s, 3H).

¹³C NMR (CD₃OD): δ=173.8, 163.7, 157.5, 152.8, 152.3, 142.4, 135.9,132.2, 132.1, 131.8, 130.7, 123.9, 122.4, 57.7, 37.7, 37.5.

Example 409 Synthesis ofN-(3-(N-Methyl-N-4-toluenesulfonylamino)pyrazin-2-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 2,3-Dichloropyrazine (Lancaster) was converted viaMethod QQQ and RRR into 2-chloro-3-nitropyrazine.L-4-(N,N-Dimethylcarbamyloxy)phenylalanine tert-butyl ester and2-chloro-3-nitropyrazine were coupled via Method BBB, and the product ofthis coupling was sequentially converted via Methods CCC, DDD, EEE andZZ to give the title compound.

Physical data were as follows:

1H NMR (CD₃OD): δ=8.07 (s, formate), 7.94 (d, 1H), 7.59 (d, 2H), 7.51(d, 1H), 7.36 (d, 2H), 7.29 (d, 2H), 7.01 (d, 2H), 4.90 (m, 1H),3.30-3.18 (m, 2H), 3.08 (s, 3H), 2.96 (s, 3H), 2.94 (s, 3H), 2.43 (s,3H).

¹³C NMR (CD₃OD): δ=177.07, 169.41, 158.64, 150.92, 147.23, 145.92,139.97, 137.14, 133.12, 129.62, 128.90, 125.69, 124.67, 124.08, 116.86,49.99, 31.67, 31.28, 30.77, 30.62, 15.46.

Example 410 Synthesis ofN-(5-(2,2,2-Trifluoroethyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 1-Trifluoromethanesulfonyloxy-2,2,2-trifluoroethanewas sequentially converted via Methods LL, OO and QQ into4,6-dichloro-5-(2,2,2-trifluoroethyl)pyrimidine.L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butyl ester and4,6-dichloro-5-(2,2,2-trifluoroethyl)pyrimidine were coupled via MethodRR, and the product of this coupling was sequentially converted viaMethods SS and ZZ to give the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.41 (s, 1H), 8.09 (s, formate), 8.06 (s, 1H), 7.24(d, 2H), 6.96 (d, 2H), 5.06 (m, 1H), 3.60-3.40 (m, 2H), 3.37-3.11 (m,2H), 3.08 (s, 3H), 2.96 (s, 3H).

¹³C NMR (CD₃OD): δ=169.35, 158.91, 156.43, 151.33, 150.97, 148.87,145.76, 130.21, 125.27, 116.80, 50.80, 31.34, 30.75, 30.60, 26.65,26.23.

Example 411 Synthesis ofN-(5-(N-Methyl-N-3-pyridinesulfonylamino)pyrimidin-4-yl)-L4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanineIsopropyl Ester

L-Tyrosine (Aldrich) was sequentially converted via Methods LLL, UU, XXand YY into L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanineisopropyl ester. 5-Nitrouracil (Aldrich) was converted via Method AAAinto 2,4-dichloro-5-nitropyrimidine.L-4-(4-Methylpiperazin-1-ylcarbonyloxy)phenylalanine isopropyl ester and2,4-dichloro-5-nitropyrimidine were coupled via Method BBB, and theproduct of this coupling was sequentially converted via Methods CCC, DDD(using 3-chlorosulfonylpyridine) and EEE to give the title compound.

Example 412 Synthesis ofN-(5-Benzylpyrimidin-4-yl)-L-4-(N,N-dimethylcarbanyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. Diethyl 2-benzylmalonate (Aldrich) was sequentiallyconverted via Methods OO and QQ into 4,6-dichloro-5-benzylpyrimidine.L-4-(N,N-Dimethylcarbamyloxy)phenylalanine tert-butyl ester and4,6-dichloro-5-benzylpyrimidine were coupled via Method RR, and theproduct of this coupling was sequentially converted via Methods SS andZZ to give the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.41 (s, 1H), 8.13 (s, formate), 7.80 (s, I H)7.34-7.19 (m, 3H), 7.17 (d, 2H), 7.00 (d, 2H), 6.85 (d, 2H), 5.01 (m,1H), 3.82 (m, 2H), 3.09 (s, 3H), 3.09-2.97 (m, 2H), 2.97 (s, 3H).

¹³C NMR (CD₃OD): δ=159.31, 156.23, 150.88, 148.07, 145.70, 141.38,131.56, 129.81, 125.30, 124.21, 124.01, 122.37, 116.81, 51.35, 31.68,30.78, 30.61, 28.28.

Example 413 Synthesis ofN-(5-(N-Methyl-N-3-pyridinesulfonylamino)pyrimidin-4-yl)-L4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalaninetert-Butyl Ester

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, XX and YY intoL-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine tert-butyl ester.5-Nitrouracil (Aldrich) was converted via Method AAA into2,4-dichloro-5-nitropyrimidine.L-4-(4-Methylpiperazin-1-ylcarbonyloxy)phenylalanine tert-butyl esterand 2,4-dichloro-5-nitropyrimidine were coupled via Method BBB, and theproduct of this coupling was sequentially converted via Methods CCC, DDD(using 3-chlorosulfonylpyridine) and EEE to give the title compound.

Example 414 Synthesis ofN-(5-(2-Trifluoromethylphenyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted via UU,VV and WW into L-4-(N,N-dimethylcarbamyloxy)phenylalanine tert-butylester. 4(3H)-Pyrimidinone (Aldrich) was sequentially converted viaMethods N and O to 4-chloro-5-iodopyrimidine.L-4-(N,N-Dimethylcarbamyloxy)phenylalanine tert-butyl ester and4-chloro-5-iodopyrimidine were coupled via Method P and the coupledproduct was reacted with 2-trifluoromethylphenyl boronic acid (Aldrich)via Method Q. The product of this coupling was converted via Method HHto give the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.51 (s, 1H), 7.84-7.49 (m, 2H), 7.71-7.63 (m, 2H),7.37 (d, 1H), 7.11-6.97 (m, 4H), 6.88 (d, 1H), 4.99 (s, 1H), 3.37-3.19(m, 1H), 3.14-3.02 (m, 4H), 2.97 (s,3H).

¹³C NMR (CD₃OD): δ=175.7, 175.5, 165.6, 161.9, 161.7, 158.6, 157.6,157.5, 153.3, 153.1, 152.6, 152.5, 136.4, 136.2, 135.0, 134.9, 134.5,133.1, 132.2, 131.9, 131.7, 128.9, 128.7, 127.8, 124.3, 123.6.

Example 415 Synthesis ofN-(5-(2-N,N-Dimethylcarbamylethyl)pyrimidin-4-yi)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 4(3H)-Pyrimidinone (Aldrich) was sequentiallyconverted via Methods N and O to 4-chloro-5-iodopyrimidine.L-4-(N,N-Dimethylcarbamyloxy)phenylalanine tert-butyl ester and4-chloro-5-iodopyrimidine were coupled via Method P and the coupledproduct was reacted with dimethylacrylamide (Aldrich) via Method U. Theproduct of this reaction which was sequentially converted via Methods Vand HH to give the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.56 (s, 1H), 8.06 (s, 1H), 7.32 (d, 2H), 7.01 (d,2H), 5.35-5.30 (m, 1H), 3.56-3.49 (m, 1H), 3.23-3.18 (m, 1H), 3.11 (s,3H), 3.02 (s, 3H), 2.99 (s, 3H), 2.97 (s, 3H), 2.88 (t, 2H), 2.65 (t,2H).

¹³C NMR (CD₃OD): δ=174.5, 174.2, 152.7, 151.6, 142.6, 136.5, 132.0,123.8, 121.0, 57.8, 38.4, 37.9, 37.5, 36.9, 32.2, 24.6.

Example 416 Synthesis ofN-(5-(N-Methyl-N-3-(1-methylpyrazole)sulfonylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanineIsopropyl Ester

L-Tyrosine (Aldrich) was sequentially converted via Methods LLL, UU, VVand WW into L-4-(N,N-dimethylcarbamyloxy)phenylalanine isopropyl ester.5-Nitrouracil (Aldrich) was converted via Method AAA into2,4-dichloro-5-nitropyrimidine. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine isopropyl ester and2,4-dichloro-5-nitropyrimidine were coupled via Method BBB, and theproduct of this coupling was sequentially converted via Methods CCC, DDD(using 1-methyl-3-chlorosulfonylpyrazole) and EEE to give the titlecompound.

Physical data were as follows:

¹H NMR (CDCl₃) δ=8.47 (s, 1H), 7.76 (s, 1H), 7.68 (bs, 2H), 7.19 (m,2H), 7.04 (d, 2H), 6.17 (d, 1H), 5.03 (m, 2H), 3.95 (s, 3H), 3.31-3.12(m, 2H), 3.08 (s, 3H), 3.06 (s, 3H), 2.99 (s, 3H), 1.24 (d, 3H), 1.21(d, 3H).

Example 417 Synthesis ofN-(6-Phenylpyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 4,6-dichloropyrimidine (Aldrich) were coupled via Method Y andthe coupled product was reacted with phenyl boronic acid (Aldrich) viaMethod Z. The product of this coupling was converted via Method HH togive the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.65 (s, 1H), 7.82-7.79 (m, 2H), 7.77-7.62 (m, 3H),7.31 (d, 2H), 7.06-7.01 (m, 4H), 5.32-5.28 (m, 1H), 3.50-3.44 (m, 1H),3.20-3.06 (m, 4H), 2.99 (s, 3H).

¹³C NMR (CD₃OD): δ=173.9, 165.7, 157.6, 154.9, 154.3, 152.8, 135.8,134.6, 132.3, 132.2, 131.7, 129.2, 123.8, 104.6, 57.8, 38.8, 37.7, 37.5.

Example 418 Synthesis ofN-(6-(2-Trifluoromethylphenyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 4,6-dichloropyrimidine (Aldrich) were coupled via Method Y andthe coupled product was reacted with 2-trifluoromethylphenyl boronicacid (Aldrich) via Method Z. The product of this coupling was convertedvia Method HH to give the title compound.

Physical data were as follows:

¹H.NMR (CD₃OD): δ=8.46 (s, 1H), 7.95-7.82 (m, 1H), 7.73-7.67 (m, 2H),7.50 - 7.48 (m, 1H), 7.29 (d, 2H), 7.03 (d, 2H), 6.65 (s, 1H), 5.05 (s,1H), 3.39 (m, 1H), 3.16-3.12 (m, 4H), 3.00 (s, 3H).

¹³C NMR (CD₃OD): δ=176.0, 164.3, 158.8, 157.7, 152.6, 136.6, 139.0,132.9, 132.1, 131.4, 130.1, 129.7, 128.2, 128.2, 123.6, 38.8, 37.7,37.5.

Example 419 Synthesis ofN-(6-(2-Hydroxymethylphenyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 4,6-dichloropyrimidine (Aldrich) were coupled via Method Y andthe coupled product was reacted with 2-(hydroxymethyl)phenyl boronicacid (Lancaster Synthesis) via Method Z. The product of this couplingwas converted via Method HH to give the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.48 (s, 1H), 8.09 (s, 1H), 7.61-7.44 (m, 4H), 7.29(d, 2H), 7.02 (d, 2H), 6.71 (s, 1H), 5.27 (s, 2H), 5.10-5.02 (m, 1H),3.42-3.41 (m, 1H), 3.16-3.12 (m, 4H), 2.99 (s, 3H).

¹³C NMR (CD₃OD): δ 175.7, 165.6, 164.7, 158.0, 157.6, 152.6, 141.6,138.5, 136.7, 135.8, 132.2, 131.9, 131.7, 131.4, 131.3, 123.7, 64.9,64.3, 38.9, 37.7, 37.5.

Example 420 Synthesis ofN-(5-Cyclohexylpyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. Cyclohexanone (Aldrich) was sequentially converted viaMethods MM, NN, OO and QQ into 4,6-dichloro-5-cyclohexylpyrimidine.L-4-(N,N-Dimethylcarbamyloxy)phenylalanine tert-butyl ester and4,6-dichloro-5-cyclohexylpyrimidine were coupled via Method RR, and theproduct of this coupling was sequentially converted via Methods SS andZZ to give the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.41 (bs, 1H), 7.89 (bs, 1H), 7.21 (d, 2H), 6.94 (d,2H), 5.12 (dd, 1H), 3.47 (dd, 1H), 3.19 (dd, 1H), 3.06 (s, 3H), 2.95 (s,3H), 3.0 (m, 1H), 2.88-2.57 (bs, 1H), 2.5 (bs, 1H), 1.95-1.67 (m, 1 H).

¹³C NMR (CD₃OD): δ=175.68, 165.82, 156.87, 152.10, 151.88, 141.96,136.30, 131.44, 125.38, 122.89, 57.86, 37.44, 36.81, 36.64, 36.30,32.65, 32.13, 27.29, 27.25, 26.95.

Example 421 Synthesis ofN-(2-(N-Methyl-N-2-furanmethylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted withN-methylfurfurylamine (Salor) via Method CC to give a product that wascoupled with o-tolyl boronic acid (Aldrich) via Method DD. The productof this coupling was converted via Method ZZ to give the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=7.43-7.35 (m, 2H), 7.35-7.2 (m, 2H), 7.2-7.0 (m, 4H),7.0-6.9 (m, 2H), 6.42 (d, 1H), 6.39 (d, 1H), 4.85 (m, 1H), 3.3-3.1 (m,7H), 3.09 (s, 3H), 2.98 (s, 3H), 2.16 (s, 3H), 1.89 (s, 3H).

Example 422 Synthesis ofN-(2-(N-Methyl-N-4-chlorophenylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted withN-methyl-4-chloroaniline (Aldrich) via Method CC to give a product thatwas coupled with o-tolyl boronic acid (Aldrich) via Method DD. Theproduct of this coupling was converted via Method ZZ to give the titlecompound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.17 (s, 1H), 7.56-7.34 (m, 8H), 7.1-6.97 (m, 4H),3.50 (m, 2H), 3.13 (s, 3H), 2.1 (s, 3H), 2.17 (s, 3H), 1.94 (s, 3H).

Example 423 Synthesis ofN-(5-(3-Thienyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 4(3H)-Pyrimidinone (Aldrich) was sequentiallyconverted via Methods N and O into 4-chloro-5-iodopyrimidine.L-4-(N,N-Dimethylcarbamyloxy)phenylalanine tert-butyl ester and4-chloro-5-iodopyrimidine were coupled via Method P and the coupledproduct was reacted with 3-thiophenyl boronic acid (Frontier Scientific,Inc.) via Method S. The product of this coupling was converted viaMethod KK to give the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.62 (s, 1H), 8.13 (s, 1H), 7.62 (m, 1H), 7.59 (m,1H), 7.20 (d, 2H), 7.09 (d, 1H), 7.01 (d, 2H), 3.47-3.13 (m, 2H), 3.13(s, 3H), 2.97 (s, 3H).

¹³C NMR (CD₃OD): δ=173.22, 162.83, 156.84, 152.17, 151.43, 141.46,135.22, 131.54, 131.35, 129.96, 127.99, 127.90, 123.24, 117.13, 56.87,36.82, 36.64.

Example 424 Synthesis ofN-(5-(2-Thienyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbarmyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 4(3H)-Pyrimidinone (Aldrich) was sequentiallyconverted via Methods N and O to 4-chloro-5-iodopyrimidine.L-4-(N,N-dimethylcarbamyloxy)phenylalanine tert-butyl ester and4-chloro-5-iodopyrimidine were coupled via Method P, and the coupledproduct was reacted with 2-thiophenyl boronic acid (Frontier Scientific,Inc.) via Method S. The product of this coupling was converted viaMethod KK to give the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.10 (s, 1H), 7.67 (s, 1H), 7.19 (d, 1H), 6.73 (m,4H), 6.49 (m, 2H), 4.80 (m, 1H), 2.89 (m, 1H), 2.70 (m, 1H), 2.60 (s,3H), 2.45 (s, 3H).

¹³C NMR (CD₃OD): δ=173.07, 162.72, 156.80, 152.13, 151.74, 142.30,135.07, 131.58, 131.14, 130.69, 130.38, 129.92, 123.19, 115.18, 56.94,36.87, 36.81, 36.62, 28.74.

Example 425 Synthesis ofN-(2-(N-Methyl-N-2-hydroxyethylamino)-5-(2-fluorophenyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted with2-(N-methylamino)ethanol (Aldrich) via Method CC to give a product thatwas coupled with 2-fluorophenyl boronic acid (Aldrich) via Method DD.The product of this coupling was converted via Method KK to give thetitle compound.

Example 426 Synthesis ofN-(5-(Piperidin-1-yl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. Piperidine (Aldrich) was sequentially converted viaMethods FFF, GGG and HHH into 4,6-dichloro-5-piperidin-1-ylpyrimidine.L-4-(N,N-Dimethylcarbamyloxy)phenylalanine tert-butyl ester and4,6-dichloro-5-piperidin-1-ylpyrimidine were coupled via Method III, andthe product of this coupling was sequentially converted via Methods JJJand ZZ into the title compound.

Example 427 Synthesis ofN-(5-(1-Propylbutyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 4-Heptanone (Aldrich) was sequentially converted viaMethods MM, NN, OO and QQ into 4,6-dichloro-5-(I-propylbutyl)pyrimidine.L-4-(N,N-Dimethylcarbamyloxy)phenylalanine tert-butyl-esterand-4,6-dichloro-5-(1-propylbutyl)pyrimidine were coupled via Method RR,and the product of this coupling was sequentially converted via MethodsSS and ZZ to give the title compound.

Example 428 Synthesis ofN-(2-(N-Methyl-N-cyclobutylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted withN-methylcyclobutylamine (prepared by the Method of Giardina et al. J.Med. Chem. 1994, 37(21), 3482-3491) via Method CC to give a product thatwas coupled with o-tolyl boronic acid (Aldrich) via Method DD. Theproduct of this coupling was converted via Method ZZ to give the titlecompound.

Example 429 Synthesis ofN-(2-(N,N-Bis-(2-hydroxyethyl)amino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

A byproduct was isolated by chromatography of the crude product ofExample 430, and the byproduct was converted via Method KK into thetitle compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=7.59 (d, 1H), 7.25 (d, 2H), 7.02 (d, 2H), 6.18 (d,1H), 3.76 (brs, 8H), 2.97 (s, 8H).

¹³C NMR (CD₃OD): δ=174.1, 163.7, 155, 152, 142.1, 135.2, 131.3, 123.7,99, 60.5, 56.8, 53.2, 37.5, 36.8, 36.6.

Example 430 Synthesis ofN-(2-(N,N-bis-(2-Hydroxyethyl)amino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted with diethanolamine(Aldrich) via Method CC to give a product that was coupled with o-tolylboronic acid (Aldrich) via Method DD. The product of this coupling wasconverted via Method KK to give the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=7.48-7.31 (m, 5H), 7.15-6.98 (m, 4H), 4.9 (m, 1H),4.63 (m, 1H), 3.83 (d, 8H), 3.1 (s, 8H), 1.9 (d, 3H).

¹³CNMR(CD₃OD): δ =173.8, 162.3, 154.6, 152.6, 140.9, 139.6, 139.4,135.9, 135.8, 132.2, 132.0, 131.4, 131.2, 131.1, 128, 123.2, 123.1,66.8, 60.6, 56.9, 56.4, 53.2, 52.8, 36.8, 36.6, 36.3, 19.5.

Example 431 Synthesis ofN-(2-(N-Methyl-N-phenylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted with N-methylaniline(Aldrich) via Method CC to give a product that was coupled with o-tolylboronic acid (Aldrich) via Method DD. The product of this coupling wasconverted via Method ZZ to give the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=7.57-6.99 (m, 14H), 4.99 (m, 1H), 3.49 (s, 3H), 3.11(m, 5H), 2.98 (s, 3H), 2.16 (s, 3H).

¹³C NMR (CD₃OD): δ=183.07, 173.72, 173.49, 162.55, 156.82, 153.97,152.07, 142.25, 141.06, 140.91, 139.53, 139.40, 135.50, 135.39, 132.21,132.16, 132.05, 131.52, 131.31, 130.53, 128.44, 128.11, 128.00, 123.13,123.04, 113.18, 56.95, 56.49, 40.02, 39.96, 37.14, 36.83, 36.65, 19.56,19.47.

Example 432 Synthesis ofN-(2-(Isopropoxy)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this coupling was sequentially converted viaMethods N,NN, DD (using o-tolyl boronic acid, Aldrich) and ZZ to givethe title compound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=7.77 (bs, 1H), 7.40-6.8 (m, 9H), 6.43 (d, 0.5H) 6.27(d, 0.5H), 6.78 (m, 1H), 6.16 (m, 1H), 3.09 (s, 3H), 3.00 (s, 3H),3.40-2.80 (m, 4H), 2.20 (s, 1.5H), 1.94 (s, 1.5H), 1.23 (m, 6H).

¹³C NMR (CDCl₃): δ=176.28, 176.15, 160.03, 159.78, 154.77, 153.65,150.01, 169.97, 139.20, 137.81, 137.64, 134.39, 134.25, 130.71, 130.47,130.12, 129.15, 126.69, 126.46, 121.24, 121.18, 109.56, 56.81, 56.34,63.19, 36.90, 36.56, 36.32, 22.19, 21.99, 21.95, 19.51, 19.27.

Example 433 Synthesis ofN-(2-(N-Methyl-N-3-methylbutylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted with N-methylN-isoamylamine (Pfaltz-Bauer) via Method CC to give a product that wascoupled with o-tolyl boronic acid (Aldrich) via Method DD. The productof this coupling was converted via Method ZZ to give the title compound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=7.6 (s, 0.5H), 7.56 (s, 0.5H), 7.30-6.80 (m, 8H) 6.30(bm, 1H), 7.00-6.00 (br, 1H), 4.63 (m, 1H), 3.09 (s, 3H), 3.01 (s, 6H),3.80-2.80 (m, 4H), 2.13 (s, 1.5H) 90 (s, 1.5H), 1.61 (m, 1H), 151 (bs,2H), 0.96 (d, 6H).

¹³C NMR (CDCl₃): δ=174.03, 173.87, 159.28, 159.04, 154.71, 153.67,150.00, 142.10, 137.81, 137.53, 134.39, 134.22, 130.78, 130.58, 130.13,128.96, 126.52, 126.30, 121.19, 121.13, 110.11, 109.91, 56.80, 56.40,48.75, 36.55, 36.33, 35.80, 25.92, 22.54, 22.48, 19.53, 19.34.

Example 434 Synthesis ofN-(2-(N-Methylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbanyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted with N-methylamine(Aldrich) via Method CC to give a product that was coupled with o-tolylboronic acid (Aldrich) via Method DD. The product of this coupling wasconverted via Method ZZ to give the title compound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=10.0-8.0 (br, 1H), 9.42 (bs, 1H), 8.24 (s, 1H),7.4-6.8 (m, 10H), 5.93 (m, 1H), 4.85 (m, 1H), 3.2-2.8 (m, 1H), 3.37 (m,1H), 3.12 (s, 1.5H), 3.11 (s, 1.5H), 3.03 (s, 1.5H), 3.02 (s, 1.5H),2.95 (s, 3H), 2.13 (s, 1.5H), 1.83 (s, 1.5H).

Example 435 Synthesis ofN-(2-(2-tolyl)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted with o-tolyl boronicacid (Aldrich) via Method KKK. The product of this coupling wasconverted via Method ZZ to give the title compound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=8.14 (d,- 1H), 7.68 (d, 1H), 7.4-6.8 (m, 12H), 5.42(m, 1H), 4.94 (m, 1H), 3.11 (s, 3H), 3.02 (s, 3H), 3.4-2.8 (m, 2H), 2.49(s, 3H), 2.11 (s, 15H), 1.91 (s, 1.5H).

Example 436 Synthesis ofN-(2-(N-Methyl-N-2-hydroxyethylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted with2-(methylamino)-ethanol (Aldrich) via Method CC to give a product thatwas coupled with o-tolyl boronic acid (Aldrich) via Method DD. Theproduct of this coupling was converted via Method ZZ to give the titlecompound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=7.4-6.94 (m, 4H), 4.82 (m, 1H), 3.8 (brs, 4H),3.23/3.26 (s, rotamers, 3H), 2.98/3.7 (s, rotamers, 6H), 1.93/2.14 (s,rotamers, 3H).

Example 437 Synthesis ofN-(2-(N-Methyl-N-2-methylpropylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted with N-methylisobutylamine (Aldrich) via Method CC to give a product that was coupledwith o-tolyl boronic acid (Aldrich) via Method DD. The product of thiscoupling was converted via Method ZZ to give the title compound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=10.5-9.8 (br, 1H), 7.63 (d, 1H), 7.3-6.8 (m, 8H),6.35(m, 1H), 4.65 (m, 1H), 3.6-2.8 (m, 4H), 3.08 (s, 3H), 3.01 (s, 6H),2.13(s, 1.5H), 2.06 (bs, 1H), 1.25 (s, 1.5H), 0.9 (s, 6H).

¹³C NMR (CDCl₃): δ=174.13, 173.97, 159.17, 158.9, 154.7, 153.99, 149.96,142.00, 137.76, 137.53, 134.50, 134.33, 130.80, 130.58, 130.15, 128.95,126.51, 126.30, 121.15, 121.11, 110.25, 109.99, 57.46, 56.90, 56.51,36.89, 36.55, 36.32, 27.08, 19.87, 19.53, 19.38.

Example 438 Synthesis ofN-(2-(N-Methyl-N-propylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted withN-methyl-N-propylamine (Aldrich) via Method CC to give a product thatwas coupled with o-tolyl boronic acid (Aldrich) via Method DD. Theproduct of this coupling was converted via Method ZZ to give the titlecompound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=10.5-9.5 (br, 1H), 7.6 (d, 1H), 7.38-6.7 (m, 8H), 6.3(m, 1H), 4.7 (m, 1H), 3.7-3.0 (m, 4H), 3.09 (s, 3H), 3.01 (s, 6H), 2.13(s, l.5H), 1.92 (s, 1.5H), 1.59 (bs, 2H), 0.89 (bs, 3H).

¹³C NMR (CDCl₃): δ=174.22, 174.06, 159.26, 159.0, 154.7, 153.76, 149.97,142.22, 137.78, 137.53, 134.53, 134.36, 130.80, 130.73, 130.51, 130.12,128.93, 126.50, 126.30, 121.16, 121.10, 110.13, 109.87, 56.90, 56.52,51.72, 36.55, 36.33, 35.96, 20.45, 19.56, 19.37, 11.06.

Example 439 Synthesis ofN-(2-(N,N-Dimethylamino)-5-(2-tolyl)pyrimidin-4-yI)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester.L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine-tert-butyl-ester and5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled via Method BB. Theproduct of this reaction was reacted with N,N-dimethylarnine (Aldrich)via Method CC to give a product that was coupled with o-tolylboronicacid (Aldrich) via Method DD. The product of this coupling was convertedvia Method ZZ to give the title compound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=11.0-9.5 (br, 1H), 7.62 (d, 1H), 7.3-6.8 (m, 8H), 6.22(m, 1H), 4.72 (m, 1H), 3.5-3.0 (m, 2H), 3.8 (s, 6H), 3.01 (s, 3H), 2.12(s, 1.5H), 1.94 (s, 1.5H).

¹³C NMR (CDCl₃): δ=174.49, 174.3, 159.4, 158.93, 154.72, 149.93, 140.30,137.75, 137.60, 134.67, 134.50, 130.92, 130.80, 130.51, 130.11, 128.87,126.48, 126.32, 121.15, 121.08, 109.87, 109.69, 56.86, 56.49, 37.51,36.87, 36.55, 36.34, 19.50, 19.38.

Example 440 Synthesis ofN-(2-(N-Methyl-N-cyclohexylamino)-5-(3-pyridyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted withN-methyl-N-cyclohexylamine (Aldrich) via Method CC to give a productthat was coupled with 3-pyridyl boronic acid 1,3-propanediol cyclicester (Lancaster Synthesis) via Method DD. The product of this couplingwas converted via Method HH to give the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.83-8.78 (m, 1H), 8.56 (brs, 1H), 8.09-7.95 (m, 2H),7.76-7.73 (m, 1H), 7.22 (d, 2H), 7.06 (d, 2H), 4.85 (m, 1H), 3.45-3.38(m, 1H), 3.18-3.11 (m, 4H), 3.06 (s, 3H), 2.99 (sm, overlapping 4H),1.92 (m, 2H), 1.76-1.57 (m, 8H).

¹³C NMR (CD₃OD): δ=173.7, 161.5, 161.4, 160.9, 157.0, 152.0, 146.0,145.7, 145.6, 143.3, 136.0, 132.2, 131.3, 128.1, 123.4, 107.8, 57.8,57.4, 36.8, 36.6, 36.1, 30.6, 30.0, 26.4, 26.2.

Example 441 Synthesis ofN-(5-(2-pheny1-2,2-difluoroethyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester.1-Trifluoromethanesulfonyloxy-2,2-difluoro-2-phenylethane wassequentially converted via Methods LL, OO and QQ into4,6-dichloro-5-(2,2-difluoro-2-phenylethyl)pyrimidine.L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butyl ester and4,6-dichloro-5-(2,2-difluoro-2-phenylethyl)pyrimidine were coupled viaMethod RR, and the product of this coupling was sequentially convertedvia Methods TT and ZZ to give the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.37 (s, 1H), 7.79 (s, 1H), 7.44 (s, 5H), 7.25 (d,2H), 6.98 (d, 2H), 5.07 (dd, 1H), 3.62-3.32 (m, 3H), 3.14 (dd, 1H) 3.08(s, 3H), 2.96 (s, 3H).

Example 442 Synthesis ofN-(5-(2-phenyl-2,2-difluoroethyl)-6-chloropyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 1-Trifluoromethanesulfonyloxy-2,2-difluoro-2-phenylethane was sequentiallyconverted via Methods LL, OO and QQ into4,6-dichloro-5-(2,2-difluoro-2-phenylethyl)pyrimidine.L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butyl ester and4,6-dichloro-5-(2,2-difluoro-2-phenylethyl)pyrimidine were coupled viaMethod RR, and the product of this coupling was converted via Method ZZto give the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.18 (s, 1H), 7.42-7.41 (m, 5H), 7.26 (d, 2H), 7.0 (d,2H), 5.03 (dd, 1H), 3.72-3.45 (m, 2H), 3.34 (dd, 1H), 3.19 (dd, 1H),3.08 (s, 3H), 2.96 (s, 3H).

Example 443 Synthesis ofN-(5-(2-phenylethyl)pyrimidin-4-yl)-L-4-(N,N-dimethylearbamyloxy)phenylalanine

4(3H)-Pyrimidinone (Aldrich) was sequentially converted via Methods Nand O into 4-chloro-5 iodopyrimidine.L-Phenylalanine-tert-butyl-ester-hydrochloride (Bachem) and4-chloro-5-iodopyrimidine were coupled via Method P. The product of thisreaction was converted via Method W to a product that was sequentiallyconverted via Methods X and HH to give the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.55 (d, 1H), 7.64 (d, 1H), 7.35-7.19 (m, 8H),7.01-6.98 (m, 2H), 5.46-5.41 (m, 1H), 5.34-3.60 (m, 1H), 3.29-3.23 (m,1H), 2.94-2.75 (m, 4H).

¹³C NMR (CD₃OD): δ=174.3, 164.3, 151.5, 141.8, 141.7, 139.2, 131.0,130.6, 130.5, 130.4, 128.9, 128.4, 120.6, 57.8, 38.4, 34.0, 30.7.

Example 444 Synthesis ofN-(2-(N-Methyl-N-cyclohexylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted withN-methyl-N-cyclohexylamine (Aldrich) via Method CC to give a productwhich was sequentially converted via Methods MMM and ZZ to give thetitle compound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=11.20 (bs, 2H), 8.44 (s, 1H), 7.76 (bs, 1H), 7.50 (br,1H), 7.18 (d, 2H), 6.96 (d, 2H), 5.91 (bs, 1H), 4.83 (bs, 1H), 4.53 (br,1H), 3.20 (m, 2H), 3.08 (s, 3H), 2.98 (s, 6H), 2.00-1.00 (m, 10H).

¹³C NMR (CDCl₃): δ=176.18, 171.50, 167.75, 162.44, 156.31, 154.49,151.52, 135.83, 131.61, 122.85, 58.04, 56.87, 38.02, 37.79, 31.16,31.00, 26.68.

Example 445 Synthesis ofN-(5-Propylpyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 4(3H)-Pyrimidinone (Aldrich) was sequentiallyconverted via Methods N and O into 4-chloro-5-iodopyrimidine.Ln4-(N,N-Dimethylcarbamyloxy)phenylalanine tert-butyl esterand-4-chloro-5-iodopyrimidine were coupled via Method P, and the productof this coupling was sequentially converted via Methods II, JJ and KK togive the title compound.

Physical data were as follows:

1H NMR (CD₃OD): δ=8.51 (s, 1H), 7.97 (s, 1H), 7.26 (d, 2H), 6.97 (d,2H), 5.36 (m, 1H), 3.51 (m, 1H), 3.23 (m, 1H), 3.16 (s, 3H), 2.95 (s,3H), 2.47 (m, 2H), 1.57 (m, 2H), 0.99 (m, 3H).

¹³C NMR (CD₃OD): δ=173.48, 163.61, 151.97, 150.75, 140.68, 135.74,133.14, 131.30, 123.02, 120.85, 56.96, 36.99, 36.76, 36.58, 29.87,21.02, 13.67.

Example 446 Synthesis ofN-(5-(2-Methoxyphenyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 4(3H)-Pyrimidinone (Aldrich) was sequentiallyconverted via Methods N and O to 4-chloro-5-iodopyrimidine.L-4-(N,N-Dimethylcarbamyloxy)phenylalanine tert-butyl ester and4-chloro-5-iodopyrimidine were coupled via Method P and the coupledproduct was reacted with 2-methoxyphenyl boronic acid (LancasterSynthesis) via Method Q. The product of this coupling was converted viaMethod HH to give the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.64 (s, 1H), 8.05 (s, 1H), 7.61-7.55 (m, 1H),7.27-7.13 (m, 5H), 6.99 (d, 2H), 5.36-5.32 (m, 1H), 3.73 (s, 3H),3.46-3.40 (m, 1H), 4.20-3.13 (m, 4H), 3.02 (s, 3H).

¹³C NMR (CD₃OD): δ=173.8, 163.5, 159.5, 157.5, 152.8, 152.1, 143.0,135.9, 134.2, 133.9, 132.2, 123.8, 123.4, 120.5, 120.0, 113.7, 57.5,57.1, 37.9, 37.7, 37.5.

Example 447 Synthesis ofN-(5-(2-Fluorophenyl)pyrimidin-4-yi)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 4(3H)-Pyrimidinone (Aldrich) was sequentiallyconverted via Methods N and O into 4-chloro-5-iodopyrimidine.L-4-(N,N-Dimethylcarbamyloxy)phenylalanine tert-butyl ester and4-chloro-5-iodopyrimidine were coupled via Method P and the coupledproduct was reacted with 2-fluorophenyl boronic acid (LancasterSynthesis) via Method Q. The product of this coupling was converted viaMethod HH to give the title compound.

Example 448 Synthesis ofN-(2-(N-Methyl-N-isopropylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted withN-methyl-N-isopropylamine (Aldrich) via Method CC to give a product thatwas coupled with o-tolyl boronic acid (Aldrich) via Method DD. Theproduct of this coupling was converted via Method ZZ to give the titlecompound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=10.5-9.5 (br, 1H), 7.59 (d, 1H), 7.30-6.70 (m, 8H),6.3 (m, 1H), 4.92 (bs, 1H), 4.7 (m, 1H), 3.50-3.0 (m, 2H), 3.08 (s, 3H),3.00 (s, 3H), 2.83 (s, 3H), 2.13 (s, 1.5H), 1.93 (s, 1.5H) 1.15 (d, 6H).

¹³C NMR (CDCl₃): δ=174.31, 174.15, 159.21, 158.95, 154.70, 153.41,149.92; 141.98, 137.79, 137.56, 134.59, 134.41, 130.59, 130.17, 128.95,126.51, 126.32, 121.15, 110.26, 110.02, 56.87, 56.50, 46.86, 36.82,36.55, 36.31, 28.18, 19.50, 19.39.

Example 449 Synthesis ofN-(2-(N-Isopropylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted with isopropylamine(Aldrich) via Method CC to give a product that was coupled with o-tolylboronicacid (Aldrich)--via Method DD. The product of this coupling wasconverted via Method ZZ to give the title compound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=9.57 (s, 1H), 8.31 (s, 1H), 7.40-6.80 (m, 8H), 6.19(m, 1H), 4.79 (m, 1H), 4.15 (m, 1H), 3.4-3.0 (m, 2H), 3.10 (s, 3H), 3.01(s, 3H), 2.16 (s, 1.5H), 1.41 (s, 1.SH), 1.24 (s, 6H).

¹³C NMR (CDCl₃): δ=176.07, 175.8, 166.23, 160.23, 159.99, 154.79,153.50, 158.06, 139.38, 137.86, 137.66, 134.10, 133.93, 130.77, 130.61,130.26, 130.01, 129.25, 126.71, 126.50, 121.46, 121.36, 109.59, 109.37,56.77, 56.22, 43.31, 36.57, 36.34, 22.12, 21.96, 19.47, 19.22.

Example 450 Synthesis ofN-(5-(2-phenylethyl)pyrimidin-4-yl)-L4-(N,N-dimethylecarbamyloxy)phenylalanineIsopropyl Ester

L-Tyrosine was sequentially converted via Methods LLL, UU, VV and WWinto L-4-(N,N-dimethylcarbamyloxy)phenylalanine isopropyl ester.4(3H)-Pyrimidinone (Aldrich) was sequentially converted via Methods Nand O to 4-chloro-5-iodopyrimidine.L-4-(N,N-Dimethylcarbamyloxy)phenylalanine isopropyl ester and4-chloro-5-iodopyrimidine were coupled via Method P, and the coupledproduct was converted via Method OOO to a product, which was convertedvia Method X to give the title compound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=8.50 (s, 1H), 7.91 (s, 1H), 7.31-7.20 (m, 3H),7.42-7.00 (m, 6H), 5.19-5.17 (m, 1H), 5.08-5.02 (m, 2H), 3.23-3.17 (m,2H), 3.06 (s, 3H), 2.99 (s, 3H), 2.83-2.78 (m, 2H), 2.65-2.60 (m, 2H),1.75-1.23 (m, 6H).

¹³C NMR (CDCl₃): δ=171.8, 159.2, 156.7, 153.5, 150.7, 140.5, 130.3,128.7, 128.5, 126.4, 121.8, 117.1, 69.4, 54.2, 36.9, 36.6, 36.5, 33.6,29.8, 21.7, 21.6.

Example 451 Synthesis ofN-(3-(N-Methyl-N-4-toluenesulfonylamino)pyrazin-2-yl)-L-phenylalanineIsopropyl Ester

L-Phenylalanine (Aldrich) was converted-via Method LLL toL-phenylalanine isopropyl ester hydrochloride. 2,3-Dichloropyrazine(Lancaster) was converted via Method QQQ and RRR into2-chloro-3-nitropyrazine. L-Phenylalanine isopropyl ester hydrochlorideand 2-chloro-3-nitropyrazine were coupled via Method BBB, and theproduct of this coupling was sequentially converted via Methods CCC, DDDand EEE to give the title compound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=7.91 (d, 1H), 7.59 (d, 2H), 7.51 (d, 1H), 7.31-7.23(m, 7H), 6.08 (d, 1H), 5.01-4.97 (m, 1H), 4.92-4.89 (m, 1H) 3.24 (d,2H), 2.97 (s, 3H), 2.43 (s, 3H), 1.21-1.12 (m, 6H).

¹³C NMR (CDCl₃): δ=167.32, 147.440, 139.85, 137.38, 133.25, 131.98,128.68, 126.17, 125.17, 125.06, 124.41, 124.11, 122.58, 64.38, 50.65,33.49, 32.41, 17.16, 17.08, 17.03.

Example 452 Synthesis ofN-(5-(2-phenylethyl)pyrimidin-4-yl)-L-phenylalanine Isopropyl Ester

L-Phenylalanine isopropyl ester hydrochloride was prepared by MethodLLL. 4(3H)-Pyrimidinone (Aldrich) was sequentially converted via MethodsN and O to 4-chloro-5-iodopyrimidine. L-Phenylalanine isopropyl esterhydrochloride and 4-chloro-5-iodopyrimidine were coupled via Method Pand the coupled product sequentially converted via Methods OOO and X togive the title compound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=8.51 (s, 1H), 7.92 (s, 1H), 7.30-7.15 (m, 5H),7.14-7.06 (m, 4H), 5.16 (m, 1H), 5.09-5.01 (m, 2H), 3.31-3.16 (m, 2H),2.79-2.74 (m, 2H), 2.62-2.57 (ni, 2H), 1.15-1.20 (m, 6H).

¹³C NMR (CDCl₃): δ=171.7, 159.1, 156.7, 153.5, 140.5, 136.1, 129.4,128.6, 128.5, 128.3, 127.1, 126.4, 117.0, 69.3, 54.2, 37.6, 33.7, 30.0,21.7, 21.6.

Example 453 Synthesis ofN-(5-(N-Methyl-N-3-pyridinesulfonylamino)pyrimidin-4-yl)-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, XX and YY intoL-4-(4-methylpiperazin-1-ylcarbonyloxy)-phenylalanine tert-butyl ester.5-Nitrouracil (Aldrich) was converted via Method AAA into2,4-dichloro-5-nitropyrimidine.L-4-(4-Methylpiperazin-1-ylcarbonyloxy)phenylalanine tert-butyl esterand 2,4-dichloro-5-nitropyrimidine were coupled via Method BBB, and theproduct of this coupling was sequentially converted via Methods CCC, DDD(using 3-chlorosulfonylpyridine), EEE and ZZ to give the title compound.

Example 454 Synthesis ofN-(2-(N-Methyl-N-cyclohexylamino)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N dimethylcarbamyloxy)-phenylalaninetert-butyl ester. L-4-(N,N-Dimethylcarbamyloxy)-phenylalanine tert-butylester and 5-bromo-2,4-dichloropyrimidine (Aldrich) were coupled viaMethod BB. The product of this reaction was reacted withN-methyl-N-cyclohexylamine (Aldrich) via Method CC to give a productthat was coupled with o-tolyl boronic acid (Aldrich) via Method DD. Theproduct of this coupling was converted via Method ZZ to give the titlecompound.

Physical data were as follows:

¹H NMR (CDCl₃): δ=10.0-9.08 (br, 1H), 7.55 (s, 0.5H), 7.52 (s, 0.5H),7.20-6.31 (m, 8H), 6.36 (br, 1H), 4.69 (m, 2H), 3.40 (m, 1H), 3.15 (m,1H), 3.06 (brs, 3H), 2.98 (brs, 3H), 2.84 (brs, 3H), 2.11 (s, 1.5H),2.00-1.00 (brm, 11.5 H).

¹³C NMR (CDCl₃): δ=164.10, 159.20, 159.00, 154.79, 153.50, 150.03,137.68, 137.48, 134.48, 130.66, 130.22, 129.01, 126.62, 126.40, 121.16,110.20, 57.00, 56.58, 55.50, 36.62, 36.39, 29.91, 29.52, 25.41, 19.60,19.65.

Example 455 Synthesis ofN-(5-(2-Tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylearbamyloxy)phenylalanineIsopropyl Ester

L-Tyrosine (Aldrich) was sequentially converted via Methods LLL, UU, VVand WW into L-4-(N,N-dimethylcarbarnyloxy)phenylalanine isopropyl ester.4(3H)-Pyrimidinone (Aldrich) was sequentially converted via Methods Nand O into 4-chloro-5-iodopyrimidine.L-4-(N,N-dimethylcarbayloxy)-phenylalanine isopropyl ester and4-chloro-5-iodopyrimidine were coupled via Method P. The product of thiscoupling was reacted with o-tolyl boronic acid via Method Q to affordthe title compound.

Physical data were as follows:

1H NMR (CDCl₃): δ=8.58 (s, 1H), 7.99 (s, 1H), 7.76-7.33 (m, 3H), 7.13(m, 0.5H), 7.03-6.95 (m, 4H), 4.97-4.87 (m, 3H), 3.08-2.99 (m, 8H), 2.09(s, 2H), 1.92 (s, 1.5H), 1.24-1.12 (m, 6H).

¹³C NMR (CDCl₃): δ=171.4, 171.2, 158.8, 158.5, 157.5, 154.7, 153.6,153.5, 150.5, 137.1, 137.0, 132.9, 132.3, 132.5, 130.8, 130.7, 130.0,129.8, 129.7, 128.9, 126.6, 126.5, 121.6, 119.5, 119.4, 69.0, 54.5,54.0, 36.9, 36.8, 36.6, 36.4, 21.65 21.60, 19.3, 19.2.

Example 456 Synthesis ofN-(5-(3-Nitrophenyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 4(3H)-pyrimidinone (Aldrich) was sequentiallyconverted via Methods N and O to 4-chloro-5-iodopyrimidine.L-4-(N,N-Dimethylcarbamyloxy)phenylalanine tert-butyl ester and4-chloro-5-iodopyrimidine were coupled via Method P, and the coupledproduct was reacted with 3-nitrophenyl boronic acid (Aldrich) via MethodT. The product of this coupling was converted via Method HH to give thetitle compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.67 (s, 1H), 8.41-8.38 (m, 1H), 8.28-8.27 (m, 1H),8.17 (s, 1H), 7.82-7.77 (m, 1H), 7.67-7.65 (m, 1H), 7.20 (d, 2H), 7.02(d, 2H), 5.33-5.28 (m, 1H), 3.47-3.411 (m, 1H), 3.12-3.04 (m, 4H), 2.97(s, 3H).

¹³C NMR (CD₃OD): δ=173.7, 163.6, 157.6, 152.8, 152.7, 151.2, 143.8,137.4, 136.3, 134.2, 133.1, 132.2, 126.7, 126.3, 124.0, 120.3, 58.0,37.7, 37.6, 37.5.

Example 457 Synthesis ofN-(5-(3-Pyridyl)pyrimidin-4-yl)-L-4-(N,N-dimethylearbamyloxy)phenylaianine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 4(3H)-Pyrimidinone (Aldrich) was sequentiallyconverted via Methods N and O to 4-chloro-5-iodopyrimidine.L-4-(N,N-Dimethylcarbamyloxy)phenylalanine tert-butyl ester and4-chloro-5-iodopyrimidine were coupled via Method P, and the coupledproduct was reacted with 3-pyridyl boronic acid 1,3-propanediol cyclicester (Lancaster Synthesis) via Method Q. The product of this couplingwas converted via Method HH to give the title compound.

Example 458 Synthesis ofN-(5-(2-phenylethyl)pyrimidin4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 4(3H)-Pyrimidinone (Aldrich) was sequentiallyconverted via Methods N and O into 4-chloro-5-iodopyrimidine.L-4-(N,N-Dimethylcarbamyloxy)phenylalanine tert-butyl ester and4-chloro-5-iodopyrimidine were coupled via Method P. The product of thisreaction was sequentially converted via Methods W, X, and HH to give thetitle compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.52 (s, 1H), 7.67 (s, 1H), 7.34-7.19 (m, 5H),7.08-6.99 (m, 4H), 5.50-5.42 (m, 1H), 5.59-5.53 (m, 1H), 3.26-3.21 (m,1H), 3.09 (s, 2H), 2.99 (s, 3H), 2.94-2.85 (m, 4H).

¹³C NMR (CD₃OD): δ=174.2, 164.2, 157.5, 152.7, 151.4, 141.8, 141.7,136.5, 132.0, 130.5, 130.4, 128.4, 123.8, 120.5, 57.8, 37.9, 37.6, 37.5,34.1, 30.6.

Example 459 Synthesis ofN-(2-N,N-Dimethylamino-5-(N-methyl-N-4-toluenesulfonylamino)pyrimidin-4-yl)-L-phenylalanine

5-Nitrouracil (Aldrich) was converted via Method AAA into2,4-dichloro-5-nitropyrimidine. L-Phenylalanine tert-butyl ester(Bachem) and 2,4-dichloro-5-nitropyrimidine were coupled via Method BBB,and the product of this coupling was sequentially converted via MethodsSSS (using dimethylamine), CCC, DDD, EEE and ZZ to give the titlecompound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.15 (s, formate), 7.65 (m, 2H), 7.41 (d, 2H),7.40-7.19 (m, 5H), 7.02-6.92 (m, 1H), 4.90 (m, 1H), 3.40-3.10 (m, 2H),3.09-2.92 (m, 9H), 2.43 (s, 3H).

¹³C NMR (CD₃OD): δ=177.07, 159.64, 154.70, 152.25, 144.10, 141.97,141.33, 140.25, 132.57, 129.02, 125.21, 124.82, 123.57, 123.42, 121.88,107.64, 51.08, 33.71, 32.72, 31.76, 15.49.

Example 460 Synthesis ofN-(5-(2-Tolyl)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

L-Tyrosine tert-butyl ester (Bachem) was sequentially converted viaMethods UU, VV and WW into L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester. 4(3H)-Pyrimidinone (Aldrich) was sequentiallyconverted via Methods N and O into 4-chloro-5-iodopyrimidine.L-4-(N,N-Dimethylcarbamyloxy)phenylalanine tert-butyl ester and4-chloro-5-iodopyrimidine were coupled via Method P and the coupledproduct was reacted with o-tolyl boronic acid (Aldrich) via Method Q.The product of this coupling was converted via Method HH to give thetitle compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ=8.75-8.65 (d, 1H), 8.05-8.03 (d, 1H), 7.51-7.35 (m, 3H), 7.26-7.11 (m, 3H), 7.02-6.97 (m, 2H), 5.38-5.27 (m, 2H), 3.50-3.39(m, 1H), 3.21-3.07 (m, 4H), 3.02 (s, 3H), 2.21-1.93 (s, 3H).

¹³C NMR (CD₃OD): δ=173.8, 173.6, 164.0, 163.8, 157.5, 152.7, 152.6,143.0, 142.8, 139.7, 139.5, 136.1, 135.9, 133.2, 133.0, 132.4, 132.2,132.1, 131.9, 131.1, 129.0, 128.9, 123.8, 123.7, 122.2, 122.0, 57.6,57.4, 37.8, 37.7, 37.5, 37.4, 20.3, 20.2.

Additionally, using the procedures described herein and the appropriatestarting materials, the following additional compounds can be prepared:

-   N-(2-(N-methyl-N-cyclohexylamino)-5-(2-methoxyphenyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine    (Example 461),-   N-(2-(N-methyl-N-isopropylamino)-5-(2-fluorophenyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine    (Example 462),-   N-(2-(N-methyl-N-isopropylamino)-5-(2-fluorophenyl)pyrimidin-4-yl)-L-4-(2-methoxyphenyl)phenylalanine    (Example 463),-   N-(2-(N-methyl-N-cyclohexylamino)-5-(2,6-difluorophenyl)pyrimidin-4-yl)-L-4-(2,6-difluorophenyl)phenylalanine    (Example 464),-   N-(2-(N-methyl-N-cyclohexylamino)-5 -(2    -hydroxymethylphenyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine    (Example 465),-   N-(2-(N,N-bis-(2-hydroxyethyl)amino)-5-(2,4,6-trimethylphenyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine    (Example 466),-   N-(2-(N-methyl-N-cyclohexylamino)-5-(2-trifluoromethylphenyl)pyrimidin-4-yl)-L-4-(2-cyanophenyl)phenylalanine    (Example 467),-   N-(2-(N-methyl-N-cyclohexylamino)-5-(3    -thienyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine    (Example 468),-   N-(2-(N-methyl-N-cyclohexylamino)-5-(2-thienyl)pyrimidin-4-yl)-L-4-(4-trifluoromethylphenyl)phenylalanine    (Example 469),-   N-(2-(N-methyl-N-cyclohexylamino)-5-(3    -pyridyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine    (Example 470),-   N-(2-(N-methyl-N-cyclohexylamino)-5-(3-nitrophenyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine    (Example 471),-   N-(2-(N-methyl-N-cyclohexylamino)-5-(2,6-dichlorophenyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine    (Example 472),-   N-(2-(N-methyl-N-cyclohexylamino)-5-(4-pyridyl)pyrimidin-4-yl)-L-4-(3-hydroxymethylphenyl)phenylalanine    (Example 473),-   N-(2-(N-ethyl-N-isopropylamino)-5-(2,6-dimethoxyphenyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine    (Example 474),-   N-(2-(N-methyl-N-cyclohexylamino)-5-(2,3    -dichlorophenyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine    (Example 475),-   N-(2-(N-methyl-N-ethylamino)-5-(2,4,6-trimethylphenyl)pyrimidin-4-yl)-L-4-(2-cyanophenyl)phenylalanine    (Example 476),-   N-(2-(N-methyl-N-isopropylamino)-5-(2,4,6-trimethylphenyl)pyrimidin-4-yl)-L-4-(3-pyridyl)phenylalanine    (Example 477),-   N-(2-(N,N-bis-(2-hydroxyethyl)amino)-5-(2,4,6-trimethylphenyl)pyrimidin-4-yl)-L-4-(2-cyanophenyl)phenylalanine    (Example 478),-   N-(2-(N-methyl-N-(1    -methylpiperidin-4-yl)amino)-5-(2-cyanophenyl)pyrimidin-4-yl)-L-4-(2,6-difluorophenyl)phenylalanine    (Example 479),-   N-(2-(N-ethyl-N-isopropylamino)-5-(2,4,6-trimethylphenyl)pyrimidin-4-yl)-L-4-(o-tolyl)phenylalanine    (Example 480),-   N-(2-(N-methyl-N-4-chlorophenylamino)-5-(2,4,6-trimethylphenyl)pyrimidin-4-′yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine    (Example 481),-   N-(5-(N-methyl-N-2-(phenyl)ethylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    (Example 482),-   N-(5-(N-methyl-N-hexylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    (Example 483),-   N-(5-(N-methyl-N-isopropylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    (Example 484),-   N-(5-(N-methyl-N-tert-butylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    (Example 485),-   N-(5-(N-ethyl-N-isopropylamino)pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    (Example 486),-   N-(5-(N-methyl-N-2-(4-pyridyl)ethyl-pyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    (Example 487),-   N-(5-(N-methyl-N-2-(phenyl)ethylamino)pyrimidin-4-yl)-L-4-(4-(2,6-dimethoxyphenyl)phenylalanine    (Example 488),-   N-(5-(N-methyl-N-hexylamino)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine    (Example 489),-   N-(5-(N-methyl-N-isopropylamino)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine    (Example 490),-   N-(5-(N-methyl-N-tert-butylamino)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine    (Example 491),-   N-(5-(N-ethyl-N-isopropylamino)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine    (Example 492),-   N-(5-(N-methyl-N-2-(4-pyridyl)ethyl-pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine    (Example 493),-   N-(2-(N-methyl-N-cyclohexylamino)-5-ethylpyrimidin-4-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine    (Example 494).

Example 495 Synthesis ofN-(4-(N,N-Di-n-hexylamino)-1,1-dioxo-1,2,5-thiadiazol-3-yl)-L-tyrosine

Step A: Preparation of 3,4-Diethyloxy-1-oxo-1,2,5-thiadiazole and3,4-Diethyloxy-1,1-dioxo-1,2,5-thiadiazole

The title intermediates were prepared according to the proceduresdescribed in R. Y. Wen et al, J Org Chem., (1975) 40, 2743; and R. Y.Wen et al, Org Prep Proceed., (1969) 1, 255.

Step B: Preparation of4-(N,N-Di-n-hexylamino)-3-ethoxy-1,1-dioxo-1,2,5-thiadiazole

Dihexylamine (90 mg, 0.48 mmol) was added to a solution of3,4-diethyloxy-1,1-dioxo-1,2,5-thiadiazole (100 mg, 0.48 mmol) inethanol (5 mL) and the reaction stirred overnight at room temperature.The solvent was removed under reduced pressure and the residue absorbedonto silica gel, and purified by flash column chromatography (silica,hexane:EtOAc 3:1) to yield the title intermediate (120 mg, 72%).

Physical data were as follows:

MS (EI, m/e) 345.

Step C: Preparation ofN-(4-(NjV-Di-n-hexylamino)-1,1-dioxo-1,2,5-thiadiazol-3-yl)-L-tyrosinetert-Butyl Ester

A solution of 4-(N,N-di-n-hexylamino)-3-ethoxy-1,1-dioxo-1,2,5-thiadiazole (400 mg, 1.02 mmol) and L-tyrosinet-butyl ester (261 mg, 1.1 mmol) in EtOH (10 mL) was stirred at roomtemperature for 36 hrs. The solvent was removed under reduced pressureresidue purified by flash column chromatography (silica, hexane:EtOAc3:1 then 1:1) to give the title compound as a white waxy solid (400 mg,73%).

Physical data were as follows:

Anal. Calcd. for C₂₇H44N₄O₅ S.0.55EtOAc: C, 59.93; H, 8.34; N, 9.57.Found: C, 59.84; H, 8.44; N, 9.62.

Step D: Preparation ofN-(4-(NiV-Di-n-hexylamino)-1,1-dioxo-1,2,5-thiadiazol-3-yl)-L-tyrosine

The compound from Step C (100 mg, 0.19 mmol) was dissolved in formicacid and the mixture stirred at room temperature for 36 hrs. Excessformic acid was removed under reduced pressure to yield the titlecompound as a white solid (90 mg, 98 %).

Physical data were as follows:

Anal. Calcd. for C₂₃H₃₆N₄O₅ S: C, 57.48; H, 7.55; N, 11.66.

Found: C, 57.04; H, 7.23; N, 11.38.

Example 496 Synthesis ofN-(4-(N,N-Di-n-hexylamino)-I,l-dioxo-1,2,5-thiadiazol-3-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Step A: Preparation ofN-(4-(N,N-Di-n-hexylamino)-1,1-dioxo-1,2,5-thiadiazol-3-yl)-L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-Butyl Ester

N-(4-(N,N-Di-n-hexylamino)-1,1 -dioxo-1,2,5-thiadiazol-3-yl)-L-tyrosinetert-butyl ester (180 mg, 0.34 mmoL) was dissolved in pyridine (5 ml).Dimethylcarbamoyl chloride (108 mg, 1 mmol) was added dropwise and themixture stirred at room temperature overnight. Pyridine was removedunder high vacuum (low water bath temperature), the residue absorbedonto silica gel and purified by flash column chromatography (silica,hexane:EtOAc 2:1) to yield the title compound (140 mg, 68%).

Step B: Preparation ofN-(4-(N,N-Di-n-hexylamino)-1,1-dioxo-1,2,5-thiadiazol-3-yl)-L-4-(N,N-dimethylcarbamyloxy)-phenylalanine

The compound from Step A (140 mg, 0.23 mmol) was dissolved in formicacid and the mixture stirred at room temperature overnight. Excessformic acid was removed under reduced pressure to yield the titlecompound as a white solid (110 mg, 87%).

Physical data were as follows:

Anal. Calcd. for C₂₆H₄₁N₅O₆ S: C, 56.6; H, 7.49; N, 12.69.

Found: C,56.67; H, 7.4; N, 12.46.

Example 497 Synthesis ofN-(4-(N,N-Di-n-hexylamino)-1,1-dioxo-1,2,5-thiadiazol-3-yl)-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine

Step A: Preparation ofN-(4-(N,N-Di-n-hexylamino)-1,1-dioxo-1,2,5-thiadiazol-3-yl)-L-4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalaninetert-Butyl Ester

A solution of N-(4-(N,N-di-n-hexylamino)-1,1 -dioxo-1,2,5-thiadiazol-3-yl)-L-ttyrosine tert-butyl ester (500 mg, 0.93 mmol), and-p-nitrophenylchloroformate (179 mg, 0.89 mmol) in dichloromethane (20 mL) was cooledto 0° C. under an argon atmosphere. Triethylamine (235 mg, 2.32 mmol)was added dropwise and the mixture stirred at 0° C. for 30 mins, thenallowed to warm to room temperature for a further 40 mins. The mixturewas recooled to 0° C. and N-methylpiperazine (90 mg, 0.89 mmol) added.The mixture was allowed to warm to room temperature and stirred forthree hours. The mixture was diluted with diethyl ether (150 mL) and theorganic solution washed with 10% potassium carbonate solution until nofurther yellow color was produced in the aqueous phase. The organiclayer was separated, dried (MgSO₄) and the solvent removed under reducedpressure. The residue was purified by flash column chromatography(silica, EtOAc:MeOH:Et₃N 94:5:1) to give the title compound as a paleyellow foam (310 mg, 50%).

Physical data were as follows:

Anal. Calcd. for C₃₃H₅₄N₆O₆ S: C, 59.79; H, 8.21; N, 12.68.

Found: C, 59.47; H, 8.25; N,12.49

Step B: Preparation ofN-(4-(N,N-Di-n-hexylainino)-1,1-dioxo-1,2,5-thiadiazol-3-yl)-L4-(4-methylpiperazin-1-ylcarbonyloxy)phenylalanine

The compound from Step A (200 mg, 0.3 mmol) was dissolved in formic acid(5 mL) and the mixture stirred at room temperature for 48 hrs. Excessformic acid was removed under reduced pressure and the residuerecrystallized from EtOAc/MeOH to yield the title compound as anoff-white solid (120 mg, 67 %).

Physical data were as follows:

Anal. Calcd. for C₂₉H₄₆N₆O₆S.0.75H₂O: C, 56.15; H, 7.72; N, 13.55.Found: C, 56.1; H, 7.44; N, 13.46.

Example 498 Synthesis ofN-[4-(2-(3-Methylphenylaminocarbonylamino)eth-1-ylamino)-1,1-dioxo-1,2,5-thiadiazol-3-yl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

Step A: Preparation of N-(4-Ethoxy-1,1-dioxo-1,2,5-thiadiazol-3-yl)L-tyrosine tert-Butyl Ester

A solution of 3,4-diethyloxy-1,1-dioxo-1,2,5-thiadiazole (400 mg, 1.94mmol)and L-tyrosine t-butyl ester (1.25 g, 5.2 mmol) in ethanol (25 mL)was stirred at room temperature overnight. Solvent was removed underreduced pressure and the product used in further transformations withoutfurther purification (Yield 790 mg).

Step B: Preparation of 2-(3-Methylphenylaminocarbonylamino)eth-1-ylamine

N-Boc-Ethylene diamine (800 mg, 5 mmol) and m-tolyl isocyanate (665 mg,5 mmol) were dissolved in acetonitrile and the mixture stirred at roomtemperature for 4 hrs. Solvent was removed under reduced pressure andthe residue absorbed onto silica gel; prior to purification by flashcolumn chromatography (silica, hexane:EtOAc 1:1) to yield the desiredcompound as a white solid (300 mg, 21%) (MS ( +ESI, m/e) 294 (M+H)⁺).The N-Boc protected compound (300 mg, 1.02 mmol) was dissolved in formicacid (10 ml) and the mixture stirred at room temperature overnight.Excess acid was removed to yield the formate salt of the title compoundas a white foam (210 mg).

Step C: Preparation ofN-[4-(2-(3-Methylphenylamino-carbonylamino)eth-1-ylamino)-1,1-dioxo-1,2,5-thiadiazol-3-yl]-L-tyrosinetert-Butyl Ester

To a solution of N-(4-ethoxy-1,1-dioxo-1,2,5-thiadiazol-3-yl)-L-tyrosinetert-butyl ester from Step A (150 mg, 0.38 mmol) and the formate salt of2-(3-methylphenylaminocarbonylamino)eth-1-ylamine from Step B (210 mg,0.89 mmol) in ethanol (10 mL) was added triethylamine (133 mg, 1.44mmol). The reaction was stirred at room temperature overnight.

Solvent was removed under reduced pressure and the residue purified byflash column chromatography (silica, 5% MeOH in EtOAc)to give the titlecompound (130 mg, 91%).

Physical data were as follows:

MS ( +ESI, m/e) 545 (M+H)⁺.

Step D: Preparation ofN-[4-(2-(3-Methylphenylamino-carbonylamino)eth-1-ylamino)-1,1-dioxo-1,2,5-thiadiazol-3-yl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine tert-Butyl Ester

The intermediate from Step C (130 mg, 0.24 mmol) was dissolved inpyridine (5 mL). Dimethylcarbamoyl chloride (77 mg, 0.72 mmol) was addeddropwise and the mixture heated at 50° C.under an argon atmosphereovernight. Pyridine was removed under reduced pressure, the residueabsorbed onto silica gel and purified by flash column chromatography(silica, hexane:EtOAc 1:2, then 5% MeOH in EtOAc) to yield the titlecompound (140 mg, 93%).

Physical data were as follows:

MS ( +ESI, m/e) 616 (M+H)⁺.

Step E: Preparation ofN-[4-(2-(3-Methylphenylamino-carbonylamino)eth-1-ylamino)-1,1-dioxo-1,2,5-thiadiazol-3-yl]-L-4-(N,N-dimethylcarbamyloxy)phenylalanine

The compound from Step D (120 mg, 0.19 mmol) was dissolved in formicacid (10 mL) and the mixture stirred at room temperature for 36 hrs.Excess acid was removed to yield the title compound as a pale yellowfoam (100 mg, 93%).

Physical data were as follows:

MS ( +ESI, m/e) 560 (M+H)⁺.

Example 499 Synthesis ofN-(4-(N,N-Dimethylamino)-1-oxo-1,2,5-thiadiazol-3-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

Step A: Preparation ofN-(4-Ethoxy-1-oxo-1,2,5-thiadiazol-3-yI)-L-tyrosine tert- Butyl Ester

A solution of 3,4-diethoxy-1-oxo-1,2,5-thiadiazole (1 g, 0.52 mmol) andL-tyrosine t-butyl ester (1.25 g, 0.52 mmol) in ethanol (25 mL) wasstirred at room temperature for 60 hr. Solvent was removed under reducedpressure and the residue purified by flash column chromatography(silica, hexane:EtOAc 1:1 to give the title intermediate (1.75 g, 88%).

Physical data were as follows:

MS ( +ESI, m/e) 382 (M+H)⁺.

Step B: Preparation ofN-(4-Ethoxy-1-oxo-1,2,5-thiadiazol-3-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

The intermediate from Step A (400 mg, 1.05 mmol) was dissolved inpyridine (10 mL) and dimethylcarbamoyl chloride (338 mg, 3.15 mmol) wasadded. The reaction was stirred at room temperature under an inertatmosphere overnight. TLC indicated large amounts of unreacted startingmaterial so the mixture was heated at 50° C. for a further 48 hrs.Excess pyridine was removed under reduced pressure and the residuepurified by flash column chromatography (silica, hexane:EtOAc 1:1 togive the title intermediate (280 mg, 59 %).

Physical data were as follows:

MS (+ESI, m/e) 453 (M+H).

Step C: Preparation ofN-(4-(N,N-Dimethylamino)-1-oxo-1,2,5-thiadiazol-3-yl)-L-4-(N,N-dimethylcarbamyloxy)phenylalaninetert-Butyl Ester

A 2M solution of dimethylamine in THF (5 mL, 10 mmol) was added to asolution of the compound from Step B (180 mg, 0.35 nmol) in ethanol (10mL). The reaction was stirred at room temperature overnight and solventremoved under reduced pressure. Residue was purified by flash columnchromatography (silica, EtOAc:MeOH:Et₃N 90:10:1) to give the titlecompound as a white foam (140 mg, 88%).

Physical data were as follows:

Anal. Caled. for C₂₂₀H₂₉N₅O₅ S: C, 53.2; H, 6.47; N, 15.51.

Found: C,52.94; H, 6.18; N,15.34.

Example 500 Synthesis ofN-(5-(2,2,2-Trifluoroethyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine

Substituting L-4-(2,6-dimethoxyphenyl)phenylalanine methyl ester fromMethod TTT for L-4-(N,N-dimethylcarbamyloxy)phenylalanine tert-butylester and following the procedure described for the preparation ofExample 410 yielded the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ 8.41 (s, 1H), 8.05 (s, 1H), 7.24 (t, 1H), 7.2 (d, 2H),7.1 (d, 2H), 6.67 (d, 2H), 5.1 (dd, 1H), 3.65 (s, 6H), 3.61-3.42 (m,2H), 3.36 (dd, 1H), 3.2 (dd, 1H).

¹³C NMR (CD₃OD): δ 175.8, 162.3, 159.2, 157.9, 155.8, 136.9, 134.4,132.2, 130.0, 129.5, 127.4, 120.9, 109.6, 105.7, 56.8, 56.2, 37.9, 32.6.

Example 501 Synthesis ofN-(2-(N-Cyclohexyl-N-methyl)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine

Substituting L-4-(2,6-dimethoxyphenyl)phenylalanine methyl ester fromMethod TTT for L-4-(N,N-dimethylcarbamyloxy)phenylalanine tert-butylester and following the procedure described for the preparation ofExample 395 yielded the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ 7.37-7.19 (m, 5.5H), 7.09-7.02 (m, 4H), 6.94 (d,0.5H), 6.68 (d, 2H), 4.79-4.74 (m, 0.5H), 4.69-4.65 (m.0.5H), 3.67 (s,3H), 3.65 (s, 3H), 3.44-3.33 (m, 1H), 3.02-2.95 (m, 4H), 2.19 (s, 1.5H),1.85-1.71 (m, 6.5H), 1.57 (m, 4H), 1.29-1.2 (br s, 1H).

Example 502 Synthesis ofN-(5-(2-Fluorophenyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine

Substituting L-4-(2,6-dimethoxyphenyl)phenylalanine methyl ester fromMethod TTT for L-4-(N,N-dimethylcarbamyloxy)phenylalanine tert-butylester and following the procedure described for the preparation ofExample 447 yielded the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ 8.50 (s, 1H), 8.01 (s, 1H), 7.3-7.0 (m, 9H), 6.69 (d,2H), 5.0 (m, 1H), 3.65 (s, 6H), 3.20-3.05 (m, 2H).

¹³C NMR (CD₃OD): δ 153.2, 151.6, 147.1, 130.2, 128.6, 126.7, 126.6,126.5, 126.4, 126.3, 123.9, 123.5, 123.2, 120.5, 120.4, 111.7, 111.4,99.6, 59.3, 31.7.

Example 503 Synthesis ofN-(2-(N-Methyl-N-propyl)-5-(2-tolyl)pyrimidin-4-yl)-L-4-(2,6-dimethoxyphenyl)phenylalanine

Substituting L-4-(2,6-dimethoxyphenyl)phenylalanine methyl ester fromMethod TTT for L-4-(N,N-dimethylcarbamyloxy)phenylalanine tert-butylester and following the procedure described for the preparation ofExample 438 yielded the title compound.

Physical data were as follows:

¹H NMR (CD₃OD): δ 10.30-8.80 (br, 1H), 7.68 (s, 0.5H), 7.63 (s, 0.5H),7.40-6.60 (m, 1H), 6.15 (m, 1H), 4.70 (m, 1H), 3.68 (s, 3H), 3.66 (s,3H), 3.80-3.00 (m, 4H), 3.07 (s, 3H), 2.12 (s, 1.5H), 2.08 (s, 1.5H),1.61 (bs, 2H), 0.87 (bs, 3H).

Example 504 Synthesis ofN-(3-chloropyrazin-2-yl)-L-4-[1-(tert-butoxycarbonyl)piperidin-4-ylcarbonylamino]phenylalanineethyl ester

Step A: Preparation of N-(3-Chloropyrazin-2-yl)-L-4-nitrophenylalanine

4-Nitrophenylalanine (50 mm, 10.59 mg) were stirred in absolute ethanolcontaining 1.0 eq (1.26 g) of sodium metal. The reaction mixture wasstripped to a brown solid and the sodium salt was taken up in 200 mL ofbutanol containing 1.0 eq (7.45 g) 2,3-dichloropyrazine. The reactionmixture was refluxed overnight and the solvent was then removed underreduced pressure. The residue was taken up in ethyl acetate and washedwith water (1×), brine (1×), dried over Na₂SO₄, filtered and stripped togive 15.5 g of the title intermediate as a brown oil.

Physical data were as follows:

Analytical: MS: (+)FAB [M+H] @ M/Z 323 with 1 Cl.

Step B: Preparation of N-(3-Chloropyrazin-2-yl)-L-4-nitrophenylalanineEthyl Ester

The intermediate from Step A was suspended in in 300 mL of absoluteethanol. The reaction flask was placed in an ice bath and cooled to 0°C. and HCl (g) was bubbled into reaction for 15 minutes. The gas tubewas replaced with a drying tube and the reaction mixture was warmed toroom temperature and stirred overnight. Ethanol was stripped off underreduced pressure to afford a dark brown residue which was taken up inethyl acetate and washed with sat. NaHCO₃ (2×), H₂O (1×), brine (1×),dried over Na₂SO₄, filtered and stripped to afford 15 g of a dark brownoil. This oil (8.0 g) was chromatographed on a silica 60 column packedin methylene chloride to provide 1.5 g (20% yield) of the titleintermediate.

Physical data were as follows:

Analytical: MS: EI M⁺ @ M/Z 350 1 Cl present.

Step C: Preparation of N-(3-Chloropyrazin-2-yl)-L-4-aminophenylalanineEthyl Ester

The intermediate from Step B (0.75 g, 0.021 mol) was placed in a Paarhydrogenation bottle with 50 mL ethanol- and 0.40 g of Pd/C catalyst.The bottle was placed on Paar shaker under 50 psi of H₂ for 3 hrs. Thereaction mixture was then fitered through a sintered glass funnel (F)and the filtered catalyst was washed with ethanol. The combinedfiltrates were stripped to a yellow oil and the oil was taken up inethyl acetate. A yellow precipitate formed and was filtered off. Thefilterate was washed with NaHCO₃ soution (1×), H₂O (1×), brine (1×),dried over Na₂SO₄, filtered and stripped to afford the titleintermediate as a yellow oil (0.340 g, 55% yield)

Step D: Preparation ofN-(3-Chloropyrazin-2-yl)-LA4-[1-(tert-butoxycarbonyl)piperidin-4-ylcarbonylamino]phenylalanineEthyl Ester

N-Boc-piperidine 4-carboxylic acid (0.253 g, 1.0 eq., 0.0011 mol) wasstirred in 30 mL methylene chloride and reaction mixture was cooled to0° C.in ice bath. HOBt (0.224 g, 1.5 eq) was added and the mixture wasstirred for 10 minutes then the intermediate from step C (1 eq., 0.32 g)was added. The reaction mixture was stirred for 5 minutes and then1,3-dicyclohexylcarbodiimide (0.25 g, 1.1 eq) was added. The reactionmixture was warmed to room temperature and stirred overnight. Thereaction was then filtered and the filtrate was stripped to give ayellow solid. The solid was taken up in ethyl acetate and filtered. Theethyl acetate solution was washed with 10% citric acid (1×), H₂O (1×),brine (1×), dried over Na₂SO₄, filtered and stripped to afford a yellowoil (0.630 g; MS: EI M⁺ @ M/Z 531 (1 chloro)). The yellow oil waschromatographed on a silica 60 colunm eluting with 3:1 hexane/ethylacetate to afford 0.997 g of the title compound. This compound may alsobe used as an intermediate for other compounds of this invention.

Physical data were as follows:

Analytical: CHN: Theory (0.5 H₂0): C, 57.71; H, 6.72; N, 12.9

Found: C, 57.79; H, 6.32; N, 12.78. MS: M⁺ @ M/Z 531 (1 Chloro).

Synthesis of Compounds of Formulae X-XV

Compounds of Formulae X-XV may be prepared as illustrated in the schemeand described in the Examples, below:

Example 505 Preparation ofN-(2-[N′-diethylamino]-5-[N″-(4-chlorophenylsulfonyl)-N″-ethylamino]pyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalanine

Step 1: Preparation of 2,4-Dichloro-5-nitropyrimidine (2).5-Nitrouracil, (1), was treated with phosphorous oxychloride (POCl₃) andN,N-dimethylaniline (PhNMe₂), according to the procedure of Whittaker(J. Chem. Soc. 195 1, 1565), to give compound 2. Compound 2 is alsoavailable from City Chemical (West Haven, Conn.).

Step 2: Preparation ofN-(2-[N′,N′-diethylaminol-5-nitropyrimidin-4-yl)-L-tyrosine tert-butylester (3). To a solution of L-tyrosine tert-butyl ester (H-Tyr(OH)-OtBu)(30.6 g, 0.129 mol) in THF (250 mL) at −10° C. was added2,4-dichloro-5-nitropyrimidine (25g, 0.129 mol), keeping the temperaturebelow 5° C.during the addition. Once the addition was complete,N,N-diisopropylethylamine (EtiPr₂N) (33.7 mL, 0.194 mol) was addeddropwise. After stirring for 1 h at −10° C., diethylamine (Et₂NH) (66.73mL, 0.645 mol) was added slowly, and then the reaction mixture waswarmed to room temperature overnight. The reaction mixture was dilutedwith diethyl ether (500 mL), and the organic layer was washed with 0.2 Ncitric acid (3×150 mL), water (1×150 mL), and 10% K₂CO₃ (3×150 mL). Theorganic phase was dried (Na₂SO₄), filtered, and concentrated in vacuo toyield a yellow residue. The residue was purified by flash chromatography(20% EtOAc/hexanes on silica gel) to yield 37.39 g (67%) of compound 3as a yellow foam. R_(f)0.21 (25% EtOAc/hexanes on silica gel).

Step 3: Preparation ofN-(2-[N′,-diethylamino]-5-nitropyrimidin4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalaninetert-butyl ester (4). To a solution ofN-(2-[N′,N′-diethylamino]-5-nitropyrimidin-4-yl)-L-tyrosine tert-butylester (37.39 g, 0.087 mol) in CH₂Cl₂ (150 mL) was added DMAP (10.59 g,0.087 mol). After 5 minutes triethylamine (TEA) (18.19 mL, 0.131 mol)was added dropwise.

1-Pyrrolidinecarbamoyl chloride (14.42 mL, 0.131 mol) was addeddropwise, and the reaction was heated to reflux (40° C.) overnight. Thereaction mixture was concentrated in vacuo and taken up in EtOAc (300mL). The organic phase was washed with 0.2 N citric acid (3×150 mL),water (1×150 mL), sat. NaHCO₃ (3×150 mL), brine (1×150 mL), dried(Na₂SO4), filtered, and concentrated in vacuo to yield 43.07 g (94%) ofcompound 4 as a yellow solid. RF 0.5 (50% EtOAc/hexanes on silica gel).

Step 4: Preparation ofN-(2-[N′,N′-diethylaminol-5-aminopyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalaninetert-butyl ester (5). A mixture ofN-(2-[N′,N′-diethylamino]-5-nitropyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalaninetert-butyl ester (43.07 g, 0.081 mol) and 10% Pd/C (4.3 g, 10 wt % Pd)in EtOH (200 mL) was shaken under 45 psi hydrogen until TLC (50%EtOAc/hexanes on silica gel) showed 100% conversion to product (48hours). The reaction mixture was then filtered through a Celite plug andconcentrated in vacuo to yield 40.29 g (100%) of compound 5 as a purplefoam. R_(f)0.11 (6:1 EtOAc/hexanes on silica gel).

Step 5: Preparation ofN-(2-[N′,N′-diethylamino]-5-[N″-(4-chlorophenyl-sulfonyl)amino]pyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalanine tert-butyl ester (6). A pyridine (160 mL) solution ofN-(2-[N′,N′-diethylamino]-5-aminopyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalaninetert-butyl ester (40.29 g, 0.081 mol) was cooled to −20° C. with a dryice/CH₃CN bath. The mixture stirred for 30 minutes, and then4-chlorobenzenesulfonyl chloride (17.06 g, 0.081 mol) was added slowly.The reaction was stirred at −20° C. to −15° C. for 4 h and then allowedto warm to room temperature overnight. The reaction was diluted withEtOAc (400 mL), and the organic phase was washed with 0.2 N citric acid(3×150 mL), water (1×150 mL), sat. NaHCO₃ (3×150 mL), brine (1×150 mL),dried (Na₂SO₄), filtered, and concentrated in vacuo to yield a brownresidue. The residue was purified by flash chromatography (50%EtOAc/hexanes on silica gel) to yield 43.49 g (80%) of compound 6 as ayellow foam. R_(f)=0.35 (50% EtOAc/hexanes on silica gel).

Step 6: Preparation ofN-(2-[N′,N′-diethylamino]-5-[N″-(4-chlorophenyl-sulfonyl)-N″-ethylamino]pyrimidin-4-yl)-4-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalaninetert-butyl ester (7). To a solution ofN-(2-[N′,N′-diethylamino]-5-[N″-(4-chlorophenyl-sulfonyl)amino]pyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalanine tert-butyl ester (42.92 g, 0.064 mol) in acetone(Me₂CO) (600 mL) was added K₂CO₃ (12.75 g, 0.096 mol), and the mixturewas stirred for 1 h at room temperature. lodoethane (EtI) (7.73 mL,0.096 mol) was then added slowly, and the reaction mixture was stirredovernight at room temperature. The reaction mixture was concentrated invacuo, and the residue was taken up in EtOAc (300 mL). The organic phasewas washed with water (2×300 mL), brine (1×100 mL), dried (Na₂SO₄),filtered, and concentrated in vacuo. The residue was purified by flashchromatography (2:1 hexanes/EtOAc on silica gel) to yield 37.36 g (85%)of compound 7 as a white solid. RF 0.53 (50% EtOAc/hexanes on silicagel).

Step 7: Preparation ofN-(2-[N′,N′-diethylamino]-5-[N″-(4-chlorophenylsulfonyl)-N″-ethylaminolpyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalaninehydrochloride (8). A formic acid (500 mL) solution ofN-(2-[N′,N′-diethylamino]-5-[N″-(4-chlorophenyl-sulfonyl)-N″ethylamino]pyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalaninetert-butyl ester (36.21 g, 0.052 mol) was heated to 70° C. for 2 h andthen concentrated in vacuo. The residue was dissolved again in formicacid (500 mL) and heated again at 70° C. for 2 h. The solution wasreduced in volume by 80% and then treated with at least 1 eq. of 1.0 NHCl (52 mL, 0.052 mol) followed by distilled water (100 mL). Theresulting heterogeneous mixture was concentrated in vacuo. Distilledwater (100 mL) was added, and the heterogeneous mixture was concentratedin vacuo. The latter steps were repeated twice to yield a wet whiteproduct. This was dried by placing under high vacuum at 40° C. (7 days)to yield 32.8 g (93%) of compound 8, as a free-flowing white solid.R_(f)=0.25 (7/3 MeOH/H₂O +0.1% TFA, reverse phase).

¹H NMR (CD₃OD) δ 8.22 (bs, 1H), 7.82-7.79 (m, 1H), 7.64-7.60 (m, 2H),7.36-7.33 (m, 1H), 7.22-7.13 (m, 2H), 7.07-6.98 (m, 2H), 4.91-4.90 (m,1H), 4.80-4.79 (m, 1H), 4.12- 4.10 (m, 1H), 3.87-3.75 (m, 1H), 3.55-3.53(m, 4H), 3.41-3.40 (m, 3H), 3.26-3.19 (m, 2H), 2.03 (bs, 1H), 1.97-1.89(m, 3H), 1.27-1.15 (m, 6H), 1.10-1.05 (t, 1.5H), 0.97-0.92 (t, 1.5H)

¹³CNMR(CD₃OD)δ 175.8, 175.7,166.5, 162.7, 162.2, 155.8, 155.7,155.7,152.6, 148.1, 147.7, 142.0, 138.5, 136.2, 132.6, 132.3, 131.9, 131.7,123.7, 111.8, 111.5, 62.3, 57.8, 44.9, 38.7, 38.0, 27.4, 26.6, 15.3,14.9, 14.7, 14.0, 13.9

Example 506 Preparation ofN-(2-[N′,N′-diethylamino]-5-[N″-(4-fluorophenylsulfonyl)-N″-ethylamino]pyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalanine

Steps 1, 2, 3, 4, 6 and 7 were performed as for Example 505. Step 5 wasperformed using 4-fluorobenzenesulfonyl chloride in place of4-chlorobenzenesulfonyl chloride.

¹H NMR (CD₃OD) δ 8.17 (bs, 1H), 7.90-7.87 (m, 2H), 7.40-7.34 (m, 2H),7.20-7.16 (m, 1H), 7.08-7.00 (m, 3H), 5.52-5.51 (m, 1H), 4.96-4.93 (m,2H), 5.78-5.70 (m, 1H), 3.85-3.75 (m, 1H), 3.59-3.53 (m, 4H), 4.47-4.43(m, 2H), 3.44-3.24 (m, 2H), 2.02-1.94 (m, 3H), 1.24-1.16 (m, 6H),1.10-1.05 (t, 1.5H), 0.99-0.94 (t, 1.5H)

¹³C NMR(CD₃OD) δ 133.0, 132.9, 132.5, 132.2, 123.7, 123.6, 118.6, 57.1,44.3, 38.3, 27.3, 26.6, 14.7, 14.1

MS m/z 629.5 (MH⁺)

Example 507 Preparation ofN-(2-([N′,N′=diethylamino]-5-[N″-(4-fluorophenysulfonyl)-N″-methylamino]pyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalanine

Steps 1, 2, 3, 4, 5 and 7 were performed as for Example 506. Step 6 wasperformed using dimethyl sulfate in place of ethyl iodide.

¹H NMR (CD₃OD) δ 8.16 (bs, 1H), 7.89-7.88 (m, 1H), 7.39-7.35 (m, 3H),7.20-7.13 (m, 1H), 7.05-7.00 (m, 2H), 4.85-4.84 (m, 1H), 4.14-4.12 (m,1H), 3.59-3.54 (m, 5H), 3.45-3.44 (m, 2H), 3.45-3.33 (m, 3H), 3.13-3.12(m, 1H), 3.02-3.01 (m, 1H), 2.04-1.95 (m, 4H), 25 1.29-1.18 (m, 6H)

¹³C NMR (CD₃OD) δ 176.5, 169.8, 166.9, 166.4, 156.2, 152.7, 151.8,150.4, 136.8, 133.3, 133.2, 132.5, 123.7, 118.8, 118.5, 57.8, 57.1,48.3, 44.5, 41.0, 38.8, 27.5, 26.7, 14.1

MS m/z 615.2 (MH+)

Example 508 Preparation ofN-(2-[N′,N′-diethylamino]-5-[N″-(4-chlorophenylsulfonyl)-N″-methylamino]pyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalanine

Steps 1, 2, 3, 4, 5 and 7 were performed as for Example 505. Step 6 wasperformed using dimethyl sulfate in place of ethyl iodide.

¹H NMR (CD₃OD) δ 8.20 (bs, 1H), 7.83-7.80 (m, 2H), 7.67-7.64 (m, 2H),7.37-7.34 (m, 1H), 7.21-7.18 (m, 1H), 7.10-7.03 (m, 2H), 4.88-4.87 (m,1H), 4.13-4.10 (m, 1H), 3.55-3.45 (m, 6H), 3.42-3.40 (m, 2H), 3.24-3.23(m, 2H), 3.11-3.10 (m, 1H), 3.02-3.01 (m, 1H), 2.04-2.03 (m, 1H),1.98-1.90 (m, 3H), 1.28-1.18 (m, 6H)

¹³C NMR (CD₃OD): δ 176.0, 166.4, 161.8, 155.9, 155.4, 152.6, 146.5,142.2, 137.6, 137.4, 136.4, 132.5, 131.9, 123.7, 114.6, 62.4, 58.1,57.7, 45.0, 40.8, 38.6, 38.3, 27.4, 26.6, 15.3, 13.9

Example 509 Preparation ofN-(2-[N′,N′-diethylamino]-5-[N′-(4-fluorophenylsulfonyl)-N″-methylamino]pyrimidin-4-yl)-41-(piperidin-1-ylcarbonyloxy)-L-phenylalanine

Steps 1, 2, 4, 5, 6 and 7 were performed as for Example 507. Step 3 wasperformed using 1-piperidinecarbonyl chloride in place of1-pyrrolidinecarbonyl chloride.

¹H NMR (CD₃OD) δ 8.16 (bs, 1H), 7.90-7.88 (m, 2H), 7.40-7.35 (m, 2H),7.21-7.20 (m, 1H), 7.14-7.13 (m, 1H), 7.02-7.01 (m, 2H), 5.51 (bs, 1H),4.83-4.77 (m, 1H), 3.64-3.53 (m, 6H), 3.34-3.33 (m, 2H), 3.20-3.17 (m,1H), 3.12-3.11 (m, 2H), 3.02-3.01 (m, 1H), 1.68-1.65 (m, 6H), 1.19-1.17(m, 6H)

¹³C NMR (CD₃OD) δ 185.0, 169.7, 166.3, 152.7, 136.6, 135.0, 133.2,133.0, 132.5, 131.8, 126.3, 123.6, 121.7, 118.6, 118.3, 57.6, 54.5,46.9, 44.3, 39.6, 38.7, 27.6, 25.9, 14.0

Example 510 Preparation ofN-(2-[N′,N′-diethylamino]-5-N″-(4-fluorophenylsulfonyl)-N″-ethylamino]pyrimidin-4-yl)-4′-(piperidin-1-ylcarbonyloxy)-L-phenylaianine

Steps 1, 2, 4, 5, 6 and 7 were performed as for Example 506. Step 3 wasperformed using 1-piperidinecarbonyl chloride in place of1-pyrrolidinecarbonyl chloride.

¹H NMR (CD₃OD) δ 8.17 (bs, 1H), 7.91-7.85 (m, 2H), 7.39-7.31 (m, 3H),7.20-7.16 (m, 1H), 7.05-6.97 (m, 2H), 4.88-4.69 (m, 2H), 4.71-4.69 (m,1H), 3.80-3.75 (m, 1H), 3.62-3.39 (m, 6H), 3.34-3.32 (m, 2H), 3.30-3.16(m, 3H), 1.68-1.65 (m, 4H), 1.23-1.17 (m, 6H), 1.10-1.05 (t, 1.5H),0.99-0.94 (t, 1.5H)

¹³C NMR (CD₃OD) δ 199.9, 187.6, 183.1, 176.2, 169.7, 166.3, 163.0,162.7, 153.9, 152.9, 136.5, 133.1, 133.0, 132.7, 132.4, 123.8, 118.8,118.4, 111.1, 110.6, 102.8, 79.4, 57.3, 55.4, 44.4, 38.9, 38.4, 27.7,26.1, 15.1, 14.8, 14.3, 14.2

Example 511 Preparation ofN-(2-[N′,N′-diethylamino]-5-[N″-(4-fluorophenylsulfonyl)-N″-ethylamino]pyrimidin-4-yl)-4′-(azetidin-1-ylcarbonyloxy)-L-phenylalanine

Steps 1, 2, 4, 5, 6 and 7 were performed as for Example 506. Step 3 wasperformed according to the following procedure.

¹H NMR (CD₃OD) δ 7.92-7.86 (m, 2H), 7.41-7.32 (m, 3H), 7.22 (d, 1H),7.04-6.91 (m, 3H), 4.29-3.98 (m, 4H), 3.88-3.72 (m, 1H), 3.69-3.37 (m,4H), 2.40-2.24 (m, 2H), 1.28- 1.11 (m, 6H), 1.10-1.00 (t, 1.5H),1.01-0.89 (t, 1.5H)

¹³C NMR (CD₃OD) δ 174.2, 169.7, 166.4, 163.2, 162.8, 157.0, 153.3,153.2, 152.4, 144.3, 143.8, 136.1, 135.6, 135.5, 133.2, 133.1, 132.5,132.2, 123.7, 118.9, 118.6, 112.9, 112.6, 57.5, 38.1, 37.7, 17.4, 14.7,14.5, 13.8, 13.7

MS m/z 615 (MH⁺)

Alternative Preparation ofN-(2-[N′,N′-diethylamino]-5-nitropyriniidin4-yl)-4′-(azetidin-1-ylcarbonyloxy)-L-phenylalaninetert-butyl ester. To a −15° C. stirred solution of compound 3 (24.9 g,0.0578 mol) and 4-nitrophenyl chloroformate (11.7 g, 0.0578 mmol) inCH₂Cl₂ (300 mL) was added triethylamine (24.2 mL, 0.173 mol), at a ratesuch that the temperature of the reaction mixture did not exceed −10° C.After stirring for 20 min, azetidine (3.30 g, 0.0578 mmol) was addeddropwise, and the reaction mixtures was warmed to room temperature andstirred overnight. The reaction mixture was diluted with EtOAc (100 mL)and hexanes (100 mL), and then was extracted repeatedly with 10% aqueousK₂CO₃, until no yellow color (4-nitrophenol) was seen in the aqueousphase. The organic layer was washed with brine (75 mL), dried withMgSO₄, filtered, and evaporated to yield 28.5 g (96%) ofN-(2-[N′,N′-diethylamino]-5-nitropyrimidin-4-yl)-4′-(azetidin-1-ylcarbonyloxy)-L-phenylalanine tert-butyl ester as a yellow solid, whichwas used without purification. R_(f)=0.17 (2:5 EtOAc/hexanes on silicagel).

Example 512 Preparation ofN-(2-[N′,N′-diethylamino]-5-[N″-(4-fluorophenylsulfonyl)-N″-methylamino]pyrimidin-4-yl)-4′-(azetidin-1-ylcarbonyloxy)-L-phenylalanine

Steps 1, 2, 3, 4, 5 and 7 were performed as for Example 511. Step 6 wasperformed using dimethyl sulfate in place of ethyl iodide.

¹H NMR (CD₃OD) δ 7.95-7.76 (m, 2H), 7.44-7.11 (m, 4H), 7.01-6.83 (m,3H), 4.30-3.93 (m, 4H), 3.66-3.41 (m, 4H), 3.14-2.92 (m, 3H), 2.42-2.21(m, 2H), 1.32-1.01 (m, 6H)

¹³H NMR (CD₃OD) δ 152.3, 136.3, 133.4, 133.2, 132.4, 123.6, 118.8,118.5, 382. 17.4, 13.8

MS m/z 601 (MH⁺)

Example 513 Preparation of N-(2-[N′,N′-diethylamino]-5-[N″-(4-chlorophenylsulfonyl)-N″-methylamino]pyrimidin-4-yl)-4′-(azetidin-1-ylcarbonyloxy)-L-phenylalanine

Steps 1, 2, 3, 4, 6 and 7 were performed as for Example 512. Step 5 wasperformed using 4-chlorobenzenesulfonyl chloride in place of4-fluorobenzenesulfonyl chloride.

¹H NMR (CD₃OD) δ 7.83 (d, 2H), 7.67 (d, 2H), 7.36-7.18 (m, 2H),7.06-6.86 (m, 3H), 4.29-3.97 (m, 4H), 3.66-3.34 (m, 5H), 3.15-2.95 (m,4H), 2.41-2.22 (m, 2H)1.26-1.06 (m, 6H)

¹³C NMR (CD₃OD) δ 157.2, 153.0, 152.5, 142.9, 142.5, 136.4, 132.5,132.1; 132.0, 123.8, 57.9, 52.2, 40.7, 38.0, 17.4, 13.6

MS m/z 617 (MH⁺)

Example 514 Preparation ofN-(2-[N′,N′-diethylamino]-5-[N″-(4-chlorophenylsulfonyl)-N′″-ethylamino]pyrimidin-4-yl)-4′-(azetidin-1-ylcarbonyloxy)-L-phenylalanine

Steps 1, 2, 3, 4, 6 and 7 were performed as for Example 511. Step 5 wasperformed using 4-chlorobenzenesulfonyl chloride in place of4-fluorobenzenesulfonyl chloride.

¹H NMR (CD₃OD) δ 7.86-7.76 (m, 2H), 7.70-7.60 (m, 2H), 7.32 (bd, 1H),7.21 (bd, 1H), 7.03-6.97 (m, 2H), 6.90 (bs, 1H), 4.29-4.00 (m, 4H),3.89-3.72 (m, 1H), 3.70-3.36 (m, 5H), 3.28-3.10 (m, 2H), 2.42-2.24 (m,2H), 1.28-1.13 (m, 6H), 1.11-1.02 (t, 1.5H), 1.01-0.90 (t, 1.5H)

MS m/z 631 (MH⁺)

Example 515 Preparation of N-(2-[N′,N′-diethylaminol-5-[N″-(2,4-difluorophenylsulfonyl)-N″-methylamino]pyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalanine

Steps 1, 2, 3, 4, 6 and 7 were performed as for Example 507. Step 5 wasperformed using 2,4-difluorobenzenesulfonyl chloride in place of4-fluorobenzenesulfonyl chloride.

¹H NMR (CDCl₃) δ 1.16 (bs, 6H), 1.93 (bs, 4H), 2.50-3.75 (m, 13H), 4.83(bs, 1H), 6.60-7.40 (m, 7H), 7.60 (bs, 1H), 7.77 (m, 1H), 9.41 (bs, 1H)

Example 516 Preparation ofN-(2-[N′,N′-diethylamino]-5-[N″-(2,4-difluorophenylsulfonyl)-N″-ethylamino]pyrimidin-4-yl)4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalanine

Steps 1, 2, 3, 4, 6 and 7 were performed as for Example 506. Step 5 wasperformed using 2,4-difluorobenzenesulfonyl chloride in place of4-fluorobenzensulfonyl chloride.

¹ H NMR (CDCl₃) δ 0.91 (t, J=6.9, 1.8H), 1.12 (m, 7.2H), 1.92 (bs, 4H),2.50-4.00 (m, 13H), 4.78 (m, 0.6H), 4.88 (m, 0.4H), 6.55 (d, J=6.9,0.4H), 6.77 (d, J=6.3, 0.6H), 6.80-7.38 (m, 6H), 7.51 (s, 0.4H), 7.58(s, 0.6H), 7.74 (m, 1H), 9.33 (m, 1H)

Example 517 Preparation ofN-(2-[N′,N′-diethylamino]-5-[N″-(2,4-difluorophenylsulfonyl)-N″-methylaminolpyrimidin-4-yl)-4′-(azetidin-1-ylcarbonyloxy)-L-phenylalanine

Steps 1, 2, 4, 5, 6 and 7 were performed as for Example 515. Step 3 wasperformed as for Example 511.

¹H NMR (CDl₃) δ 1.14 (t, J=6:6, 6H), 2.32 (m, 2H), 2.50-3.80 (m, 9H),4.13 (m, 4H), 4.62 (m, 0.6H), 4.81 (m, 0.4H), 5.81 (bd, 0.6H), 5.90 (bd,0.4H), 6.90-7.40 (m, 7H), 7.77 (m, 1 H)

MS m/z 619.2 (MH⁺)

Example 518 Preparation ofN-(2-N′,N′-diethylamino]-5-[N″-(2,4-difluorophenylsulfonyl)-N″-ethylamino]pyrimidin-4yl)4′-(azetidin-1-ylcarbonyloxy)-L-phenylalanine

Steps 1, 2, 4, 5, 6 and 7 were performed as for Example 516. Step 3 wasperformed as for Example 511.

¹H NMR (CDCl₃) δ 0.89 (t, J=6.7, 1.8H), 1.16 (m, 7.2H), 2.28 (m, 2H),3.00-4.00 (m, 8H),.4.09 (bs, 4H), 4.79 (m, 0.6H), 4.88 (m, 0.4H),6.80-7.30 (m, 7H), 7.57 (s, 0.4H), 7.62 (s, 0.6H), 7.75 (m, 1H), 11.9(bs, 1H)

MS m/z 633.2 (MH⁺)

Example 519 Preparation ofN-(2-N′,N′-diethylamino]-5-[N″-(4-fluorophenylsulfonyl)-N″-propargylaminolpyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalanine

Steps 1, 2, 3, 4, 5 and 7 were performed as for Example 506. Step 6 wasperformed using propargyl bromide in place of ethyl iodide.

¹H NMR (CDC′₃) δ 1.18 (m, 6H), 1.93 (bs, 4H), 2.37 (s, 1H), 3.00-3.70(m, 10H), 3.80 (d, J=21.3, 0.6H), 3.98 (d, J=18.3, 0.4H), 4.51 (m, 1H),4.88 (m, 11H), 6.75-7.35 (m, 7H), 7.58 (s, 0.6H), 7.63 (s, 0.4H), 7.86(m, 2H), 9.71 (bs, 1H)

Example 520 Preparation ofN-(2-[N′,N′-diethylamino]-5-[N″-(2,4-difluorophenylsulfonyl)-N″-propargylamino]pyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalanine

Steps 1, 2, 3, 4, 5 and 7 were performed as for Example 515. Step 6 wasperformed using propargyl bromide in place of dimethyl sulfate.

¹H NMR (CDCl₃) δ 1.17 (m, 6H), 1.94 (m, 4H), 2.40 (m, 1H), 3.00-3.75 (m,10H), 3.99 (d, J=18.0, 0.6H), 4.18 (d, J 18.0, 0.4H), 4.50 (m, 1H), 4.90(m, 1H), 6;75-7.35 (m, 7H), 7.81 (m, 2H), 10.0 (bs, 1H)

Example 521 Preparation ofN-(2-[N′,N′-diethylamino]-5-[N″-(2,4-difluorophenylsulfonyl)-N″-propargylamino]pyrimidin-4-yl)-4′-(azetidin-1-ylcarbonyloxy)-L-phenylalanine

Steps 1, 2, 4, 5, 6 and 7 were performed as for Example 520. Step 3 wasperformed as for Example 511.

¹H NMR (CDCl₃) δ 1.18 (m, 6H), 2.34 (m, 31H), 3.00-3.75 (m, 6H),3.80-4.25 (m, 5H), 4.47 (m, 1H), 4.89 (m, 1H), 6.75-7.35 (m, 7H), 7.79(m, 2H), 10.3 (bs, 1H)

MS m/z 643.2 (MH⁺)

Example 522 Preparation ofN-(2-[N′,N′-diethylamino]-5-[N″-(4-fluorophenylsulfonyl)-N″-propargylamino]pyrimidin-4-yl)-4′-(azetidin-1-ylcarbonyloxy)-L-phenylalanine

Steps 1, 2, 3, 4, 5 and 7 were performed as for Example 511. Step 6 wasperformed using propargyl bromide in place of ethyl iodide.

¹H NMR (CDCl₃) δ 1.25 (m, 6H), 2.28 (m, 3H), 3.00-3.75 (m, 6H),3.80-4.25 (m, 5H), 4.47 (m, 1H), 4.89 (m, 1H), 6.75-7.35 (m, 7H), 7.57(s, 0.6H), 7.62 (s, 0.4H), 7.79 (m, 2H), 10.6 (bs, 1H)

MS m/z 625.2 (MH⁺)

Example 523 Preparation ofN-(2-N′,N′-diethylamino]-5-[N″-(4-chlorophenylsulfonyl)-N″-propargylamino]pyrimidin-4-yl)-4′-(pyrrolidin-1-ylcarbonyloxy)-L-phenylalanine

Steps 1, 2, 3, 4, 5 and 7 were performed as for Example 505. Step 6 wasperformed using propargyl bromide in place of ethyl iodide.

¹H NMR (CD₃OD) δ 8.13 (s, 1H), 7.86-7.82 (m, 2H), 7.62-7.58 (m, 2H),7.32-7.28 (m, 2H), 7.19-7.17 (m, 1H), 7.04-6.98 (m, 2H), 4.83-4.5(m,-2H), 4.12-3.82 (m, 1H), 3.63--3.37.(m,8H),-3.27-3.08 (m, 2H), 2.72(bs, 1H), 2.04-1.86 (m, 4H), 1.24-1.07 (m, 6H)

¹³C NMR (CD₃OD) δ 177.2, 176.5, 162.7, 156.7, 155.7, 154.5, 153.2,142.6, 140.3, 137.4, 137.3, 133.1, 132.9, 132.8, 132.7, 132.2, 132.1,124.3, 111.3, 80.5, 80.3, 77.7, 58.2, 57.7, 44.9, 43.4, 28.1, 27.3,14.8, 14.7

MS m/z 655 (MH⁺)

Synthesis of Compounds of Formulae XVI-XXI

Compounds of Formulae XVI-XXI may be prepared as described in theExamples below:

Example 524 Preparation of2-{2-diethylamino-5-[(4-fluorobenzenesulfonyl)methylamino]pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic acid

General. Flash chromatography was performed using a Biotage Flash 75L,using 800 g KP-Sil silica cartridges (32-63 μM, 60 angstrom, 500-550m²/g). Ris are reported for analytical thin layer chromatography, usingEM Science Silica Gel F(254) 250 μM thick plates for normal phase, andWatman MKC18F 200 μM thick plates for reverse phase.

Step 1: Preparation of 2,4-Dichloro-5-nitropyrimidine. 5-Nitrouracil,was treated with phosphorous oxychloride and N,N-dimethylaniline,according to the procedure of Whittaker (J. Chem. Soc. 1951, 1565), togive the title compound, which is also available from City Chemical(West Haven, Conn.).

Step 2: Preparation of2-(2-diethylamino-5-nitropyrimidin-4-ylamino)-3-(4-hydroxyphenyl)propionicacid, t-butyl ester. To absolution of2-amino-3-(4-hydroxyphenyl)propionic acid, (30.6 g, 0.129 mol) in THF(250 mL) at -1 0C was added 2,4-Dichloro-5-nitropyrimidine (25g, 0.129mol), keeping the temperature below 5° C. during the addition. Once theaddition was complete, N,N-diisopropylethylamine (33.7 mL, 0.194 mol)was added dropwise. After stirring for 1 h at −10° C., diethylamine(66.73 mL, 0.645 mol) was added slowly, and then the reaction mixturewas warmed to room temperature overnight. The reaction mixture wasdiluted with diethyl ether (500 mL), and the organic layer was washedwith 0.2 N citric acid (3×150 mL), water (1×150 mL), and 10% K₂CO₃(3×150 mL). The organic phase was dried (Na₂SO₄), filtered,- andconcentrated in vacuo to yield a yellow residue. The residue waspurified by flash chromatography (20% EtOAc/hexanes on silica gel) toyield 37.39 g (67%) the title compound as a yellow foam. R_(f)=0.21 (25%EtOAc/hexanes on silica gel).

Step 3: Preparation of2-(2-diethylamino-5-nitropyrimidin-4-ylamino)-3-(4-dimethylcarbamoyloxyphenyl)propionicacid t-butyl ester. To a solution of2-(2-diethylamino-5-nitropyrimidin-4-ylamino)-3-(4-hydroxy-phenyl)propionicacid t-butyl ester (31.80 g, 0.074 mol) in CH₂Cl₂ (600 mL) was addedDMAP (9.00 g, 0.074 mol). After 5 minutes triethylamine (10.23 mL, 0.074mol) was added dropwise. N,N-dimethylcarbamyl chloride (13.83 mL, 0.110mol) was added dropwise, and the reaction was heated to refluxovernight. The reaction mixture was concentrated in vacuo and taken upin EtOAc (1 L). The organic phase was washed with 0.5 M citric acid(3×250 mL), sat. NaHCO₃ (3×250 mL), brine (1×250 mL), dried (MgSO₄),filtered, and concentrated in vacuo to yield 37.0 g (99%) the titlecompound as a white solid.

Step 4: Preparation of2-(2-diethylamino-5-aminopyrimidin4-ylamino)-3-(4-dimethylcarbamoyloxyphenyl)propionicacid t-butyl ester. A mixture of2-(2-diethylamino-5-nitropyrimidin-4-ylamino)-3-(4-dimethylcarbamoyl-oxyphenyl)propionic acid t-butyl ester(37.0 g,0.073 mol) and 10% Pd/C (3.8 g, 10 wt% Pd) in EtOH (250 mL) was shakenunder 60 psi hydrogen until TLC (50% EtOAc/hexanes on silica gel) showed100% conversion to product (48 hours). The reaction mixture was thenfiltered through a Celite plug and concentrated in vacuo to yield 32.0 g(92%) the title compound as a violet foam.

Step 5: Preparation of 2-{2-diethylamino-5-[(4-fluorobenzenesulfonyl)amino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl) propionicacid t-butyl ester. A pyridine (120 mL) solution of2-(2-diethylamino-5-aminopyrimidin-4-ylamino)-3-(4-dimethylcarbamoyloxy-phenyl)propionicacid t-butyl ester(32.0 g, 0.067 mol) was cooled to −20° C. with a dryice/CH₃CN bath. The mixture stirred for 30 minutes, and thenp-fluorobenzenesulfonyl chloride (13.18 g, 0.067 mol) was added slowly.The reaction was stirred at −20° C. for 4.5 hrs, and then3-dimethylaminopropyl amine (8.52 mL, 0.067 mol) was added, and then themixture was allowed to warm to room temperature overnight. The reactionwas concentrated in vacuo. The residue was taken up in EtOAc (1 L), andthe organic phase was washed with 0.5 M citric acid (3×900 mL), water(1×900 mL), sat. NaHCO₃ (3×900 mL), brine (1×900 mL), dried (MgSO₄),filtered, and concentrated in vacuo to yield a brown residue. Theresidue was purified by flash chromatography (50% EtOAc/hexanes onsilica.gel) to yield 33.04 g (77%) the title compound as a yellow foam.R_(f)=0.54 (3:2 EtOAc/hexanes on silica gel).

Step 6: Preparation of 2-{2-diethylamino-5-[(4-fluorobenzenesulfonyl)methylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic acid t-butyl ester. To a solution of2-{2-diethylamino-5-[(4-fluorobenzenesulfonyl)amino]-pyrimidin-4-ylamino}-3-(4-dimethyl-carbamoyloxyphenyl)propionicacid t-butyl ester (33.04 g, 0.052 mol) in acetone (510 mL) was addedK₂CO₃ (8.69 g, 0.063 mol), and the mixture was stirred for 10 min atroom temperature. Dimethyl sulfate (5.95 mL, 0.063 mol) was then addedslowly, and the reaction mixture was stirred overnight at roomtemperature. The reaction mixture was concentrated in vacuo, and theresidue was taken up in EtOAc (600 mL). The organic phase was washedwith water (2×400 mL), brine (2×400 mL), dried MgSO₄, filtered, andconcentrated in vacuo. The residue was purified by flash chromatography(2:1 hexanes/EtOAc on silica gel) to yield 28.69 g (85%) the titlecompound as a white solid.

Step 7: Preparation of 2-{2-diethylamino-5-[(4-fluorobenzenesulfonyl)methylamino]pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic acid hydrochloride. A formic acid (500 mL) solution of 2-{2-diethylamino-5-[(4-fluorobenzenesulfonyl)methylamino]-pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic acid t-butyl ester (28.69 g, 0.044 mol) was heated to 70° C.for 2 h, and then concentrated in vacuo. The residue was dissolved againin formic acid (500 mL), and then heated again at 70° C. for 2 h, andthen concentrated again in vacuo. The residue was dissolved again informic acid (500 mL), and then heated again at 70° C. for 1 h. Thesolution was reduced in volume by 90%, and then treated with 1.0 M HCl(44 mL, 0.044 mol) and distilled water (490 mL). The resultinghomogeneous solution was concentrated in vacuo, and then distilled water(100 mL) was added, and the homogenous solution was lyophilized over 14days to yield 26.76 g (96%) the title compound, as a white solid.

¹H NMR (CD₃OD) δ 7.96-7.92 (m, 2H), 7.45-7.25 (m, 4H), 7.06-6.95 (m,3H), 5.00-4.93 (m, 1H), 3.55-3.40 (m, 5H), 3.34-3.20 (m,, 2H), 3.15-3.05(m, 5H), 3.07-3.00 (m, 3H), 1.22 (bs, 6H)

¹³CNMR (CD₃OD) δ 171.6, 168.3, 154.5, 144.4, 137.9, 135.1, 135.0, 134.1,125.5, 120.6, 120.3, 39.6, 39.2, 39.1, 15.2

MS m/z 589 (MH+)

Example 525 Preparation of2-{2-diethylamino-5-[(4-chlorobenzenesulfonyl)methylamino]pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic acid

Steps 1, 2, 3, 4, 6 and 7 were performed as for Example 524. Step 5 wasperformed using 4-chlorobenzenesulfonyl chloride in place of4-fluorobenzenesulfonyl chloride.

¹H NMR (CD₃OD) δ 7.88-7.85 (m, 2H), 7.72-7.69 (m, 2H), 7.39-7.25 (m,2H), 7.14- 6.92 (m, 3H), 5.00-4.85 (m, 1H), 3.60-3.50 (m, 1H), 3.37-3.28(m, 6H), 3.15-3.07 (m, 6H), 3.01 (bs, 3H), 1.22 (bs, 6H)

¹³C NMR (CD₃OD) δ 208.6, 145.3, 134.9, 128.8, 124.9, 124.5, 124.4,116.3, 50.2, 30.4, 30.0, 6.0

MS m/z 605 (MH⁺)

Example 526 Preparation of2-{2-diethylamino-5-[(3,4-difluorobenzenesulfonyl)methylamino]pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic acid

Steps 1, 2, 3, 4, 6 and 7 were performed as for Example 524.Step-5 wasperformed using 3,4-difluorobenzenesulfonyl chloride in place of4-fluorobenzenesulfonyl chloride.

¹H NMR (CD₃OD) δ 7.84-7.77 (m, 1H), 7.67 (bs, 1H), 7.58-7.53 (m, 1H),7.37-7.34 (m, 1H), 7.22-7.18 (m, 1H), 7.08-7.02 (m, 3H), 4.83-4.76 (m,1H), 3.55-3.54 (m, 4H), 3.35-3.33 (m, 1H), 3.23-3.12 (m, 6H), 3.03-2.99(m, 3H), 1.19 (bs, 6H)

¹³C NMR (CD₃OD)δ 178.3, 177.8, 163.2, 162.6, 159.3, 159.1, 155.9, 155.7,154.3, 153.0, 152.5, 152.4, 138.4, 138.1, 134.0, 129.5, 125.3, 122.4,122.2, 121.7, 121.4, 115.3, 59.3, 46.0, 42.4, 41.9, 40.4, 39.9, 39.2,39.1, 15.76

MS m/z 607.2 (MH+)

Example 527 Preparation of2-{2-diethylamino-5-[(3,4-dichlorobenzenesulfonyl) methylanminolpyrimidin4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl) propionic acid

Steps 1, 2, 3, 4, 6 and 7 were performed as for Example 524. Step 5 wasperformed using 3,4-dichlorobenzenesulfonyl chloride in place of4-fluorobenzenesulfonyl chloride.

¹H NMR (CD₃OD) δ 8.00-7.98 (m, 1H), 7.83-7.74 (m, 2H), 7.37-7.34 (m,1H), 7.21- 7.20 (m, 1H), 7.10-7.02 (m, 3H), 4.85-4.83 (m, 1H), 3.55-3.53(m, 2H), 3.35-3.33 (m, 1H), 3.21-3.12 (m, 6H), 3.04-2.99 (m,.6H), 1.19(bs, 6H)

¹³C NMR (CD₃OD) δ 176.4, 166.2, 161.7, 161.2, 158.0, 157.8, 152.8,151.5, 150.5, 140.2, 139.8, 139.5, 136.8, 135.8, 133.9, 132.6, 132.0,129.8, 123.8, 113.7, 113.4, 57.8, 44.6, 40.8, 40.4, 38.7, 38.3, 37.7,37.5, 14.1

MS m/z 639.1 (MH+)

Example 528 Preparation of2-{2-diethylamino-5-[(benzenesulfonyl)methylamino]pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic acid

Steps 1, 2, 3, 4, 6 and 7 were performed as for Example 524. Step 5 wasperformed using benzenesulfonyl chloride in place of4-fluorobenzenesulfonyl chloride.

¹H NMR (CD₃OD) δ 8.14 (bs, 1H), (7.85-7.84 (m, 1H), 7.8-7.78 (m, 1H),7.69-7.66 (m,-2H), 7.40-7.37- (m-, 1 H), 7.21-7.195 (m, 1H). 7.04-7.03(m, 2H), 7.95-7.90 (m,-1H), 5.52 (bs, 1H), 3.54-3.53 (m, 2H), 3.36-3.33(m, 6H), 3.13-3.12 (m, 3H), 3.01-3.00 (m, 3H), 1.20-1.17 (m, 6H)

¹³C NMR (CD₃OD) δ 165.9, 152.8, 136.7, 135.8, 132.6, 131.6, 130.2,123.8, 44.7, 37.5, 14.0

MS m/z 571.2 (MH+)

Example 529 Preparation of2-{2-diethylamino-5-[(2-fluorobenzenesulfonyl)methylamino]pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic acid

Steps 1, 2, 3, 4, 6 and 7 were performed as for Example 524. Step 5 wasperformed using 2-fluorobenzenesulfonyl chloride in place of4-fluorobenzenesulfonyl chloride.

¹H NMR (CD₃OD) δ 8.31 (bs, 1H), 7.94-7.85 (m, 2H), 7.57-7.44 (m, 3H),7.34-7.30 (m, 1H), 7.15-7.12 (m, 2H), 5.00-4.85 (1H), 3.63-3.62 (m, 4H),3.50-3.42 (m, 1H), 3.34-3.30 (m, 4H), 3.29-3.22 (m, 4H), 3.11-3.10 (m,2Hi), 1.28 (bs, 6H)

¹³C NMR (CD₃OD) δ 176.5, 166.4, 163.1, 160.4, 159.7, 157.7, 152.8,151.5, 150.7, 138.5, 138.3, 136.7, 133.7, 132.5, 132.2, 127.1, 123.7,119.9, 119.6, 113.4, 57.8, 44.6, 40.6, 39.0, 38.4, 37.7, 37.5, 14.1

Example 530 Preparation of2-{2-diethylamino-5-[(3-fluorobenzenesulfonyl)methylamino]pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic acid

Steps 1, 2, 3, 4, 6 and 7 were performed as for Example 524. Step 5 wasperformed using 3-fluorobenzenesulfonyl chloride in place of4-fluorobenzenesulfonyl chloride.

¹H NMR (CD₃OD) δ 8.15-8.12 (bs, 1H), 7.72-7.68 (m, 1H), 7.63-7.60 (m,1H), 7.53-7.52 (m, 1H), 7.38-7.35 (m, 1H), 7.21-7.20 (m, 1H), 7.10-6.99(m, 3H), 4.87-4.86 (m, 1H), 3.54-3.53 (m, 4H), 3.35-3.34 (m, 3H),3.15-3.12 (m, 4H), 3.05-3.00 (m, 4H), 1.20 (bs, 6H)

¹³C NMR (CD₃OD) δ 166.1, 153.1, 136.9, 134.1, 132.8, 126.5, 124.1,123.2, 122.9, 117.7, 117.4, 103.4, 45.0, 38.0, 14.3

MS m/z 589.2 (MH+)

Example 531 Preparation of2-{2-diethylamino-5-[(4-fluorobenzenesulfonyl) isopropylaminolpyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl) propionic acid

Steps 1, 2 and 3 were performed as for Example 524. Thereafter, Steps 4and 6 were accomplished in one pot, according to the followingprocedure. Thereafter, Steps 5 and 7 were performed as for Example 524.

¹H NMR (CD₃OD) δ 8.20-8.16 (m, 1H), 7.95-7.84 (m, 2H), 7.36-7.25 (m,3H), 7.24-7.15 (m, 3H), 7.07-6.98 (m, 3H), 5.07-5.05 (m, 1H), 4.90-4.86(m, 1H), 4.65-4.62 (m, 1H), 4.49-4.41 (m, 1H), 3.63-3.56 (m, 3H),3.38-3.31 (m, 2H), 3.27-3.11 (m, 2H), 3.00-2.99 (m, 3H), 1.27-1.21 (m,6H), 1.05-0.99 (m, 6H)

¹³C NMR (CD₃OD) δ 175.8, 175.5, 169.6, 166.3, 165.9, 163.5, 163.4,157.7, 153.0, 152.9, 152.3, 138.1, 136.4, 136.1, 133.1, 133.0, 133.0,132.9, 132.7, 132.3, 123.8, 118.8, 118.7, 118.5, 118.4, 107.5, 57.6,57.2, 54.7, 44.7, 38.7, 38.1, 37.6, 37.5, 23.0, 22.9, 22.2, 22.0, 14.1,14.0

Alternative one-pot procedure for the preparation of2-(2-diethylamino-5-isopropylaminopyrimidin4-yl)-3-(4-dimethylcarbamoyloxyphenyl)propionic acid t-butyl ester. A mixture of2-(2-diethylamino-5-nitropyrimidin-4-ylamino)-3-(4-dimethylcarbamoyloxyphenyl)propionicacid t-butyl ester (5.0 g, 0.010 mol), glacial acetic acid (10 drops),acetone (2.19 mL, 0.030 mol), and platinum oxide (0.250 g, 5 wt %) inEtOH (15 mL) was hydrogenated at 45 psi hydrogen until TLC (50%EtOAc/hexanes) showed 100% conversion to product (20 hours). Thereaction mixture was then filtered through a Celite plug andconcentrated in vacuo to yield a brown residue. The residue was purifiedby flash chromatography (4:1 EtOAc/hexanes) to yield 3.54 g (70%) 9 as apurple foam.

Example 532 Preparation of2-{2-diethylamino-5-1(4-fluorobenzenesulfonyl)ethylaminolpyrimidin4-ylamino}-3-(4- dimethylcarbamoyloxyphenyl)propionic acid

Steps 1, 2, 3, 4, 5 and 7 were performed as for Example 524. Step 6 wasperformed using ethyl iodide in place of dimethyl sulfate.

¹H NMR (CDCl₃) δ 0.89 (t, J=7.2, 1.8H), 1.06 (t, J=7.1, 1.2H), 1.10-1.30(m, 6H), 2.97 (s, 3H), 3.05 (s, 3H), 3.10-3.90 (m, 8H), 4.82 (q; J=5.4,0.6H), 4.91 (q, J=6.1, 0.4H), 6.80-7.45 (m, 8H), 7.77 (m, 2H), 12.44(bs, 1H)

MS m/z 603.3 (MH⁺)

Example 533 Preparation of2-{2-diethylamino-5-[(3,4-difluorobenzenesulfonyl)isopropylaminolpyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic acid

Steps 1, 2, 3, 4, 6 and 7 were performed as for Example 531. Step 5 wasperformed using 3,4-difluorobenzenesulfonyl chloride in place of4-fluorobenzenesulfonyl chloride.

¹H NMR (CD₃OD) δ 8.20-8.19 (m, 1H), 7.84-7.78 (m, 1H), 7.70-7.64 (m,1H), 7.54- 7.48 (m, 1H), 7.39-7.31 (m, 1H), 7.20-7.17 (m, 1H), 7.05-6.96(m, 2H), 4.91-4.89 (m, 1H), 4.70-4.68 (m, 1H), 4.48-4.41 (m, 2H),3.60-3.58 (m, 3H), 3.34-3.33 (m, 1H), 3.27-3.20 (m, 1H), 3.09-3.08 (m,2H), 2.98-2.97 (m, 2H), 1.28-1.19 (m, 6H), 1.06-0.98 (m, 6H), 0.83-0.81(m, 1H)

¹³C NMR(CD₃OD)δ 177.6, 177.2, 167.9,164.9, 164.8, 159.2, 159.1, 155.7,154.5, 154.4, 152.4, 152.3, 140.4, 140.3, 137.8, 134.3, 133.9, 129.3,129.2, 125.4, 122.6, 122.5, 122.4, 122.2, 121.5, 121.2, 109.1, 59.5,59.1, 56.7, 56.6, 46.4, 46.3, 39.6, 39.3, 39.2, 24.7, 24.5, 23.9, 23.6,15.7, 15.6

Example 534 Preparation of 2-{2-diethylamino-5-[(4-chlorobenzenesulfonyl) isopropylaminolpyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl) propionic acid

Steps 1, 2, 3, 4, 6 and 7 were performed as for Example 531. Step 5 wasperformed using 4-chlorobenzenesulfonyl chloride in place of4-fluorobenzenesulfonyl chloride.

¹H NMR (CD₃OD) 8 8.18-8.17 (m, 1H), 7.85-7.78 (m, 1H), 7.62-7.58 (m,1H), 7.38- 7.35 (m, 1H), 7.34-7.24 (m, 1H), 7.17-7.16 (m, 1H), 7.10-7.05(m, 2H), 7.04-6.98 (m, 2H), 4.98-4.87 (m, 1H), 4.73-4.68 (m, 1H),4.55-4.38 (m, 2H), 3.70-3.52 (m, 3H), 3.40-3.30 (m, 1H), 3.28-3.18 (m,1H), 3.17-3.08 (m, 2H), 3.05-2.98 (m, 2H), 1.25-1.20 (m, 6H), 1.04-0.96(m, 6H), 0.80-0.77 (m, 1H)

¹³C NMR (CD₃OD) δ 175.7, 175.5, 166.2, 165.8, 169.6, 163.5, 163.4,157.6, 152.9, 152.8, 138.0, 136.3, 136.1, 133.1, 133.0, 132.9, 132.7,132.2, 123.8, 118.8, 118.6, 118.5, 118.5, 118.3, 107.5, 57.6, 57.2,54.7, 44.6, 38.6, 38.1, 37.6, 37.5, 22.9, 22.8, 22.2, 21.9, 14.1, 13.9

Example 535 Preparation of2-{2-diethylamino-5-[(3,4-difluorobenzenesulfonyl)ethylaminolpyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic acid

Steps 1, 2, 3, 4, 6 and 7 were performed as for Example 532. Step 5 wasperformed using 3,4-difluorobenzensulfonyl chloride in place of4-fluorobenzenesulfonyl chloride.

¹H NMR (CD₃OD) δ 8.15-8.14 (m, 1H), 7.80-7.75 (m, 1H), 7.73-7.62 (m,1H), 7.60-7.49 (m, 1H), 7.30-7.18 (m, 1H), 7.16-7.00 (m, 2H), 5.58-5.50(m, 1H), 4.90-4.83 (m, 1H), 5.78-5.70 (m, 1H), 3.85-3.75 (m, 1H),3.65-3.54 (m, 3H), 3.40-3.23 (m, 5H), 3.18-3.10 (m, 3H), 3.05-2.98 (m,3H), 1.25-1.15 (m, 3H), 1.18-1.05 (t, 1.5H), 1.02-1.00 (t, 1.5H)

¹³C NMR (CD₃OD) δ 165.8, 152.7,145.7, 136.4, 136.3, 132.5, 132.2, 127.5,123.6, 120.7, 120.4, 81.4, 57.0, 44.3, 38.5, 38.1, 37.4, 14.9, 14.6,14.1, 14.0

MS m/z 621.5 (MH+)

Example 536 Preparation of2-{2-diethylamino-5-[(4-chlorobenzenesulfonyl)ethylaminolpyrimidin4-ylamino}-3-(4- dimethylcarbamoyloxyphenyl)propionic acid

Steps 1, 2, 3, 4, 6 and 7 were performed as for Example 532. Step 5 wasperformed using 4-chlorobenzenensulfonyl chloride in place of4-fluorobenzenesulfonyl chloride.

¹H NMR (CD₃OD) δ 8.15-8.14 (m, 1H), 7.84-7.79 (m, 1H), 7.67-7.61 (m,1H), 7.37-7.33 (m, 1H), 7.22-7.18 (m, 1H), 7.14-7.13 (m, 1H), 7.06-7.00(m, 3H), 4.80-4.75 (m, 1H), 4.18-4.10 (m, 1H), 3.65-3.30 (m, 3H),3.28-3.20 (m, 3H), 3.18-3.08 (m, 2H), 3.03-2.98 (m, 2H), 2.05-2.04 (m,1H), 1.30-1.16 (m, 9H), 1.10-1.08 (t, 1.5H), 0.99-0.95 (t, 1.5H)

¹³CNMR(CD₃OD)δ 176.2, 176.1, 166.7, 162.7, 162.3, 157.6, 152.9, 142.0,138.8, 136.5, 132.8, 132.5, 132.0, 131.8, 123.8, 111.7, 111.4, 57.9,57.8, 44.9, 38.9, 38.3, 37.8, 37.7, 15.1, 14.9, 14.3, 14.2

MS m/z 619.4 (MH+)

Example 537 Preparation of2-{2-diethylamino-5-[(4-fluorobenzenesulfonyl)cylclopropylmethylamino]pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyl-oxyphenyl)propionicacid

Steps 1, 2, 3, 4, 5 and 7 were performed as for Example 524. Step 6 wasperformed using bromomethylcyclopropane and cesium carbonate in place ofdimethyl sulfate and potassium carbonate.

¹H NMR (CDCl₃) δ 0.2-0.2 (m, 2.4H), 0.2-0.45 (m, 1.6H), 0.54 (m, 0.6H),0.85 (m, 0.4H), 1.00-1.40 (m, 6H), 2.80-3.80 (m, 14H), 4.79 (q; J=5.5,0.6H), 4.91 (q, J=6.3, 0.4H), 6.70-7.40 (m, 8H), 7.77 (m, 2H), 10.26(bs, 1H)

MS m/z 629.2 (MH⁺)

Example 538 Preparation of2-{2-diethylamino-5-1(3,5-difluorobenzenesulfonyl)methylamino]pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxypheyl)propionic acid

Steps 1, 2, 3, 4, 6 and 7 were performed as for Example 524. Step 5 wasperformed using 3,5-difluorobenzenesulfonyl chloride in place of4-fluorobenzenesulfonyl chloride.

¹H NMR (CD₃OD) δ 7.68-7.67 (m, 1H), 7.67-7.56 (m, 2H), 7.42-7.40 (m,2H), 7.31- 7.30 (m, 1H), 7.26-7.23 (m, 2H), 5.20-4.90 (m, 1H), 4.35-4.33(m, 1H), 3.78-3.74 (m, 4H), 3.57-3.54 (2H), 3.38-3.33 (m, 2H), 3.26-3.21(m, 2H), 2.41-2.39 (m, 2H), 2.26-2.25 (m, 2H), 4.50-1.38 (m, 6H)

¹³C NMR (CD₃OD) δ 162.5, 162.3, 159.2, 159.0, 148.0, 146.1, 132.2,127.8, 127.7, 127.6, 118.9, 109.1, 109.0, 108.7, 108.6, 106.2, 105.8,52.5, 39.6, 34.1, 32.9, 9.5

Example 539 Preparation of2-{2-diethylamino-5-[(3,5-difluorobenzenesulfonyl)ethylamino]pyrimidin4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic acid

Steps 1,2, 3, 4, 5 and 7 were performed as for Example 538. Step 6 wasperformed using ethyl iodide in place of dimethyl sulfate.

¹H NMR (CD₃OD) δ 7.45-7.43 (m, 1H), 7.42-7.18 (m, 2H), 7.21-7.16 (m,2H), 7.07-7.06 (m, 1H), 7.04-6.97 (m, 2H), 5.51 (bs, 1H), 4.86-4.82 (m,1H), 4.72-4.66 (m, 1H), 3.84-3.77 (m, 1H), 3.59-3.50 (m, 3H), 3.34-3.31(m, 2H), 3.12-3.10 (m, 3H), 2.99-2.96 (m, 3H), 1.22-1.14 (m, 9H),1.10-1.05 (t, 1.5H), 0.97-0.95 (t, 1.5H)

¹³C NMR (CD₃OD) δ 159.9, 150.9, 150.1, 134.0, 130.0, 129.7, 121.2,107.9, 86.7, 42.0, 41.9, 36.3, 35.2, 35.1, 12.8, 12.5, 11.9, 11.8,

Example 540 Preparation of2-{2-diethylamino-5-[(2,4-difluorobenzenesulfonyl)methylamino]pyrimidin4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic acid

Steps 1, 2, 3, 4, 6 and 7 were performed as for Example 524. Step 5 wasperformed using 2,4-difluorobenzenesulfonyl chloride in place of4-fluorobenzenesulfonyl chloride.

¹H NMR (CD₃OD) δ 8.16-8.11 (m, 1H), 7.59-7.56 (m, 2H), 7.48-7.45 (m,2H), 7.26-7.24 (m, 3H), 5.21-5.16 (m, 1H), 3.79-3.77 (m, 4H), 3.57-3.54(m, 3H), 3.48-3.46 (m, 2H), 3.44-3.34 (m, 3H); 3.22-3.21 (m, 3H),1.45-1.44 (m, 6H)

¹³C NMR (CDCl) δ 180.2, 170.3, 166.6, 150.3, 129.0, 128.9, 128.7, 125.9,125.4, 117.5, 117.4, 116.5, 114.8, 107.7, 107.4, 95.5, 90.8, 68.0, 65.1,55.7, 50.8, 37.6, 36.4, 31.9, 31.7, 31.6, 13.2, 9.4, 8.3, 7.8

Example 541 Preparation of2-{2-diethylamino-5-[(2,4-difluorobenzenesulfonyl) ethylamino]pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl) propionic acid

Steps 1, 2, 3, 4, 5 and 7 were performed as for Example 540. Step 6 wasperformed using ethyl iodide in place of dimethyl sulfate.

¹H NMR (CD₃OD) δ 8.15 (bs, 1H), 7.91-7.76 (m, 1H), 7.32-7.30 (m, 2H),7.20-7.19 (m, 2H), 7.04-7.00 (m, 2H), 4.84-4.83 (m, 1H), 4.74-4.67 (m,1H), 4.14-4.07 (m, 1H), 3.92-3.82 (mi 1H), 3.51-3.49 (mn, 3H), 3.34-3.31(m, 3H), 3.12-2.99 (m, 2H), 2.98-2.97 (m, 2H), 2.03-2.02 (m, 1H),1.26-1.17 (m, 6H), 1.10-1.06 (t, 1.5H), 1.03-0.98 (t, 1.5H)

¹³C NMR (CD₃OD) δ 173.6, 173.3, 171.4, 167.7, 164.3, 161.2, 159.9,159.3, 157.1, 156.7, 155.2, 152.4, 151.0, 150.3, 134.0, 133.3, 133.1,132.9, 130.0, 123.2, 122.9, 122.8, 121.3, 121.2, 112.0, 111.8, 111.6,111.5, 107.7, 107.2, 106.0, 105.9, 105.6, 105.2, 60.0, 54.8, 42.0, 36.5,35.9, 35.3, 35.1, 19.3, 13.0, 12.9, 12.7, 11.9, 11.8

Example 542 Preparation of2-{2-diethylamino-5-[(3,5-dichlorobenzenesulfonyl) methylamino]pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl) propionic acid

Steps 1, 2, 3, 4, 6 and 7 were performed as for Example 524. Step 5 wasperformed using 3,5-dichlorobenzenesulfonyl chloride in place of4-fluorobenzenesulfonyl chloride.

¹H NMR (CD₃OD) δ 7.84-7.82 (m, 1H), 7.76-7.75 (m, 3H), 7.34-7.32 (m,1H), 7.19-7.10 (m, 1H), 7.03-7.00 (m, 2H), 5.50 (bs, 1 H), 4.83-4.82 (m,1 H), 4.74-7.73 (m, 1H),3.55-3.38 (m, 4H), 3.34-3.32 (m, 2H), 3.15-3.11(m, 4H), 3.02-2.99 (m, 3H), 1.18-1.15 (m, 6H

¹³C NMR (CD₃OD) δ 157.1, 155.2, 150.1, 149.7, 140.1, 135.9, 134.3,132.9, 130.0, 129.9, 126.0, 121.2, 110.7, 55.2, 54.8, 42.0, 38.5, 38.1,36.5, 35.9, 35.2, 35.1, 11.9

MS m/z 639.1 (MH+)

Example 543 Preparation of2-{2-diethylamino-5-[(3,5-dichlorobenzenesulfonyl) ethylamino]pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl) propionic acid

Steps 1, 2, 3, 4, 5 and 7 were performed as for Example 542. Step 6 wasperformed using ethyl iodide in place of dimethyl sulfate.

¹H NMR (CD₃OD) δ 8.15 (bs, 1H), 7.84-7.84-7.79 (m, 1H), 7.76-7.74 (m,2H), 7.33-7.30 (m, 1H), 7.22-7.11 (m, 2H), 7.04-6.98 (m, 1H), 5.51 (bs,1H), 4.86-4.82 (m, 1H), 4.72-4.67 (m, 1H), 3.77-3.75 (m, 1H), 3.60-3.50(m, 3H), 3.34-3.29 (m, 2H), 3.27-3.22 (m, 2H), 3.12-3.11 (m, 2H),2.99-2.98 (m, 2H), 1.23-1.14 (m, 6H), 1.10-1.05 (t, 1.5H), 0.99-0.94 (t,1.5H)

¹³CNMR(CD₃OD)δ 173.6, 173.4, 163.7, 159.9, 159.3, 157.3, 156.8, 155.2,155.1, 152.1, 150.8, 150.2, 141.4, 141.2, 135.9, 134.0, 132.7, 130.0,129.7, 125.8, 125.7, 121.3, 121.2, 107.9, 107.4, 54.8, 54.7, 42.0, 36.4,35.8, 35.3, 35.1, 12.8, 12.5, 11.9, 11.8

MS m/z 653.2 (MH+)

Example 544 Preparation of2-{2-diethylamino-5-[(4-fluorobenzenesulfonyl)-n-propylamino]pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl) propionic acid

Steps 1, 2, 3, 4, 5 and 7 were performed as for Example 524. Step 6 wasperformed using 1-propyl iodide in place of dimethyl sulfate.

¹H NMR (CDCl₃) δ 0.75 (m, 3H), 1.00-1.50 (m, 8H), 3.00 (s, 3H), 3.08 (s,3H), 3.20-3.70 (m, 8H), 4.79 (q, J=6.3, 0.6H), 4.91 (q, J=6.6, 0.4H),5.73 (bs, 0.6H), 5.92 (bs, 0.4H), 6.90-7.45 (m, 7H), 7.76 (m, 2H)

MS m/z 617.2 (MH⁺)

Example 545 Preparation of2-{2-diethylamino-5-1(4-fluorobenzenesulfonyl)allylamino]pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic acid

Steps 1, 2, 3, 4, 5 and 7 were performed as for Example 524. Step 6 wasperformed using allyl bromide in place of dimethyl sulfate.

¹H NMR (CDCl₃) δ 1.20 (m, 6H), 2.98 (s, 3H), 3.06 (s, 3H), 3.10-4.30 (m,8H), 4.75-4.95 (m, 1H), 5.07 (m, 2H), 5.48 (m,.0.6H), 5.67 (m, 0.4H),6.90-7.45 (m, 8H), 7.76 (m, 2H), 11.07 (bs, 1H)

MS m/z 615.2 (MH⁺)

Example 546 Preparation of2-{2-diethylamino-5-[(4-fluorobenzenesulfonyl)isobotylamino]pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic acid

Steps 1, 2, 3, 4, 5 and 7 were performed as for Example 524. Step 6 wasperformed using isobutyl iodide in place of dimethyl sulfate.

MS m/z 631.2 (MH⁺)

Example 547 Preparation of2-{2-diethylamino-5-[(4-fluorobenzenesulfonyl)-n-butylamino]pyrimidin4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionicacid

Steps 1, 2, 3, 4, 5 and 7 were performed as for Example 524. Step 6 wasperformed using 1-butyl iodide in place of dimethyl sulfate.

¹H NMR (CDCl₃) δ 0.82 (q, J=7.1, 3H), 1.05-1.40 (m, 10H), 3.01 (s, 3H),3.10 (s, 3H), 3.15-3.80 (m, 8H), 4.75 (q, J=6.3, 0.6H), 4.91 (q, J=5.9,0.4H), 5.79 (d, J=5.4, 0.6H), 5.91 (d, J=6.6, 0.4H), 7.00-7.40 (m, 7H),7.77 (m, 2H)

Example 548 Preparation of2-12-diethylamino-5-[(2,5-difluorobenzenesulfonyl) methylamino]pyrimidin-4-ylamino}3-(4-dimethylcarbamoyloxyphenyl) propionic acid

Steps 1, 2, 3, 4, 6 and 7 were performed as for Example 524. Step 5 wasperformed using 2,6-difluorobenzenesulfonyl chloride in place of4-fluorobenzenesulfonyl chloride.

¹H NMR (CD₃OD) δ 8.38-8.37 (m, 1H), 7.99-7.95 (m, 1H), 7.55-7.54 (m,2H), 7.50-7.42 (m, 2H), 7.27-7.22 (m, 2H), 5.08-5.06 (m, 1H), 3.76-3.74(m, 4H), 3.59-3.54 (m, 3H), 3.49-3.42 (m, 4H), 3.36-3.34 (m, 2H),3.23-3.21 (m, 2H), 1.40 (bs, 6H)

¹³C NMR (CD₃OD) δ 161.4, 159.2, 155.8, 153.1, 148.1, 147.1, 133.6,132.0, 127.8, 119.0, 111.1, 110.8, 110.7, 108.5, 105.8, 94.8, 86.4,66.7, 54.0, 52.8, 39.7, 35.8, 34.2, 33.7, 32.9, 32.8, 9.4

Example 549 Preparation of2-{2-diethylamino-5-[(2,3-difluorobenzenesulfonyl) ethylamino]pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl) propionic acid

Steps 1, 2, 3, 4, 6 and 7 were performed as for Example 524. Step 5 wasperformed using 2,3-difluorobenzenesulfonyl chloride in place of4-fluorobenzenesulfonyl chloride. 2,3-Difluorobenzenesulfonyl chloridewas prepared by the following procedure.

1H NMR (CD₃OD) δ 8.32 (bs, 1H), 7.90-7.80 (m, 2H), 7.59-7.48 (m, 3H),7.27-7.23 (m, 2H), 5.09-5.08 (m, 1H), 3.77-3.70 (m, 4H), 3.60-3.51 (m,3H), 3.50-3.42 (m, 2H), 3.39-3.3 In (m, 3H), 3.32-3.18 (m, 2H),1.43-1.41 (m, 6H)

¹³C NMR (CD₃OD) δ 170.4, 160.8, 158.1, 156.1, 153.0, 151.6, 150.5,148.9, 148.2, 147.3, 147.2, 143.9, 143.5, 142.6, 141.1, 140.9, 131.8,127.7, 125.1, 123.8, 120.8, 120.6, 119.2, 40.5, 35.7, 33.4, 32.9, 32.7,9.0

Preparation of 2,3-Difluorobenzenesulfonyl Chloride. The followingprocedure was executed using two flasks. In the first flask,2,3-difluoroaniline (2.0 g, 0.015 mol). was dissolved in concentratedHCl (15.9 mL), and the resulting solution was cooled to −5° C., using anice/NaCl bath. A solution of sodium nitrite (1.18 g, 0.017 mol) indistilled water (13.6 mL) was added in portions with stirring, whilemaintaining the temperature below 0° C., and the mixture was stirred for10 min. In the second flask, thionyl chloride (5.08 mL, 0.069 mol) wasadded dropwise to distilled water (30.6 mL), which had been pre-cooledto −5° C., using an ice/NaCi bath. The resulting solution was allowed towarm to room temperature, and then Cu(I)Cl (0.08 g, 0.77 mmol) wasadded, and then the reaction mixture was re-cooled to −5° C. Withcontinued cooling and stirring, the contents of the first flask wereadded in 2 mL portions to the contents of the second flask, and themixture was stirred for 30 min, during which time a precipitate formed.The precipitate was isolated by filtration, rinsed with cold water, andstored under vacuum to give 3.25 g (98%) 10 as a white solid.

Example 550 Preparation of2-{2-Diethylamino-5-[(4-fluorobenzenesulfonyl)propargylamino]pyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic acid

Steps 1, 2, 3, 4, 5 and 7 were performed as for Example 524. Step 6 wasperformed using propargyl bromide in place of dimethyl sulfate.

¹H NMR (CDCl₃) δ 1.15 (m, 6H), 2.27 (d, J=2.1, I H), 2.97 (s, 3H), 3.06(s, 3H), 3.10-3.70 (m, 6H), 3.75 (dd, J=17.7, 2.0, 0.6H), 3.95 (dd,J=18.1, 2.0, 0.4H), 4.51 (dd, J=19.5, 22, 0.6H), 4.54 (dd, J=18.1, 2.2,0.4H), 4.79 (q, J=5.9, 0.6H), 4.88 (q, J=6.6, 0.4H), 6.42 (bd, 0.4H),6.65 (bs, 0.6H), 6.85-7.30 (m, 6H), 7.52 (s, 0.6H), 7.56 (s, 0.4H), 7.85(m, 2H), 8.20 (bs, 1H)

MS m/z 613.2 (MH⁺)

Example 551 Preparation of2-{2-Diethylamino-5-[(2,4-difluorobenzenesulfonyl)propargylaminolpyrimidin-4-ylamino}-3-(4-dimethylcarbamoyloxyphenyl)propionic acid

Steps 1, 2, 3, 4, 5 and 7 were performed as for Example 540. Step 6 wasperformed using propargyl bromide in place of dimethyl sulfate.

¹H NMR (CDCl₃) δ 1.16 (q, J=7.5, 6H), 2.27 (m, 1H), 2.99 (s, 3H), 3.09(s, 3H), 3.10-3.70 (m, 6H), 4.04 (dd, J=17.7, 2.4, 0.6H), 4.24 (dd,J=17.9, 2.2, 0.4H), 4.47 (m, 1H), 4.81 (q, J=5.9, 0.6H), 4.89 (q, J=6.3,0.4H), 6.27 (d, J=7.5, 0.4H), 6.41 (d, J=5.7, 0.6H), 6.90-7.10 (m, 4H),7.16 (d, J=8.3, 1H), 7.28 (d, J=8.3, 1H), 7.55 (bs, 1H), 7.66 (s, 0.6H),7.67 (s, 0.4H), 7.81 (m, 1H)

Example 552 Preparation of2-{2-diethylamino-5-[(4-fluorobenzenesulfonyl)-(2,2,2-trifluoroethyl)aminolpyrimidin-4-ylamino}-3-(4-dimethylcarbamoyl-oxyphenyl)propionic acid

Steps 1, 2, 3, 4, 5 and 7 were performed as for Example 524. Step 6 wasperformed using 2,2,2-trifluoroethyl triflate and cesium carbonate inplace of dimethyl sulfate and potassium carbonate.

¹H NMR (CDCl₃) δ 1.14 (m, 6H), 2.98 (s, 3H), 3.06 (s, 3H), 3.10-4.20 (m,8H), 4.80 (q, J=5.9, 0.6H), 4.87 (q, J=6.2, 0.4H), 6.09 (d, J=5.9,0.4H), 6.18 (bd, 0.6H), 6.80-7.50 (m, 7H), 7.55 (bs, 1H), 7.77 (m, 2H)

MS m/z 657.2 (MH+)

Synthesis of Compounds of PEG Derivatives

The following methods may be used to prepare the compounds of thisinvention. In one method outlined in Scheme 16 below is illustrative ofsuch preparation.

The following Examples describe methods for preparing the compoundsshown in Scheme 6 and Scheme 16 above. Unless otherwise indicated someor all of the following HPLC methods were used in the preparation of thefollowing exemplary compounds.

Method A1: Samples of conjugates of more than 100 mg were purified usingreverse phase HPLC on a Phenomenex Luna C18(2), 5 μm column 250 mm×21.2mm with a Varian UV detector, using a gradient of 40-60% ACN+0.1% TFA in100 min at 15 mL/min.

Method B1: Samples of conjugates of more than 100 mg but less than 500mg were purified using reverse phase HPLC on a Phenomenex Luna C18(2),10 μm column 250 mm×50 mm with a Varian UV detector using a gradient of40-60% ACN +0.1% TFA in 100 min at 60 mL/min.

Method C1: The purity of conjugates was confirmed using reverse phaseHPLC on a Luna 3 μm C18(2) column (30×4.6 mm) with a Sedex 75 (35° C.,gain=5) evaporative light scattering detector, using a gradient of20-70% ACN w/0.1% TFA at a flow rate of 1.5 mL/min.

Example 553 Preparation of 2 kDa urea-linked mPEG conjugate carboxylicacid

Step 1: Preparation of compound 29

Compound 25 (20 g, 0.11 mol) (as shown in Scheme δabove) was dissolvedin CH₂Cl₂ (500 mL) under N₂. The reaction mixture was cooled to 0° C..Triethylamine (18.12 mL, 0.13 mol) was added, followed bytrifluoroacetic anhydride (18.14 mL, 0.13 mol) in portions. The reactionwas allowed to warm to room temperature overnight. The reaction mixturewas concentrated in vacuo and the residue was taken up in ethyl acetate(200 mL). The organic phase was washed with H₂O, sat. NaHCO₃, brine,dried over Na₂SO₄, filtered, and concentrated in vacuo to yield 29.73 g(96%) of the title compound, 29, as a yellow solid.

¹H NMR (CDCl₃) δ 3.64-3.60 (m, 2H), 3.55-3.53 (m, 2H), 3.49-3.45 (m,4H), 1.44 (s, 9H).

¹³C NMR (CDCl₃) δ 155.7 (J_(C-F)=36 Hz), 154.3, 116.4 (J_(C-F)=288 Hz),80.8, 45.7, 43.3, 28.3.Step 2: Preparation of compound 30

Compound 29 (29.26 g, 0.10 mol) was added in portions to a 500 mL flaskcontaining a solution of 4N HCl in dioxane (200 mL) at 0° C. Thereaction was stirred in ice bath for 4 hours when TLC (3:1 hexanes:ethyl acetate) showed 100% conversion to product. The reaction mixturewas concentrated in vacuo and treated with ethyl ether (500 mL). Theproduct was filtered and dried to yield 22.53 g (99%) compound 30 as awhite mono-hydrochloride salt.

¹H NMR (DMSO-d₆) δ 3.82-3.79 (m, 4H), 3.53 (s, 1H), 3.18-3.16 (m, 4H).

¹³C NMR (DMSO-d₆) δ 154.3 (J_(C-F)=35 Hz), 115.9 (J_(C-F=)289 Hz), 66.1,42.0, 41.9, 41.5.Step 3: Preparation of compound 31

A 250 mL flask was charged with compound 30 (1.0 g, 4.6 mmol), CH₂Cl₂(40 mL), and sat. NaHCO₃ (40 mL). The reaction mixture was stirredvigorously at 0° C. for 15 minutes. Stirring was ceased and the layerswere allowed to separate. A 2.0 M solution of phosgene in toluene (9 mL,18 mmol) was added to the reaction mixture which was stirred vigorouslyfor 30 minutes, while maintaining temperature at 0° C. The layers wereseparated and the aqueous phase was washed with CH₂Cl₂ (15 mL). Thecombined organic layers were washed with brine, dried over Na₂SO₄,filtered, and concentrated in vacuo. The residue was taken up in CH₂Cl₂and concentrated in vacuo again to yield 1.04 g (92%) compound 31 as awhite solid.

MS(PI-FAB) 245, (M+H)⁺.

¹H NMR (CDCl₃) δ 3.80-3.68 (m, 8H).

¹³C NMR (CDCl₃) δ 155.9 (J_(C-F)=37 Hz), 148.7 (J_(C-F)=12 Hz), 116.3(J_(C-F)=289 Hz), 48.3, 47.8, 45.7, 45.3, 45.1, 42.9, 42.7.Step 4: Preparation of compound 32

A 25 mL flask was charged with compound 24 (5.97 g, 0.011 mol), DMAP(1.34 g, 0.011 mol), and CH₂Cl₂ (22 mL). Triethylamine (2.4 mL, 0.017mol) was added followed by compound 31 (4.2 g, 0.017 mol). The reactionmixture was heated at reflux for 20 hours. The reaction mixture wasconcentrated in vacuo and the residue was taken up in ethyl acetate. Theorganic phase was washed with sat. NaHCO₃, H₂O, brine, dried overNa₂SO₄, filtered, and concentrated in vacuo to yield 9.31 g (115%) pinkfoam. The crude material was purified by flash chromatography (gradientof 50% ethyl acetate/hexanes to 75% ethyl acetate/hexanes) to yield 6.1g (76%) compound 32 as apale pink foam. R_(f)=0.14 (1:1 hexanes:ethylacetate).

MS(PI-FAB) 730, (M+H)⁺.

¹H NMR (CDCl₃) δ 9.08-9.07 (m, 1H), 8.87-8.85 (m, 1H), 8.16-8.14 (m,1H), 7.52-7.48 (m, 1H), 7.25-7.22 (d, 2H), 7.03-7.00 (d, 2H), 6.91-6.88(d, 1H), 4.78-4.70 (q, 1H), 4.60-4.44 (dd, 2H), 3.88 (s, 1H), 3.75-3.60(m, 8H), 3.09-3.06 (m, 2H), 1.42 (s, 9H), 1.18 (s, 3H), 1.16 (s, 3H).Step 5: Preparation of compound 33

To a solution of compound 32 (6.11 g, 8.4 mmol) dissolved in MeOH (90mL) was added a solution of potassium carbonate (5.79 g, 42 mmol) in H₂O(10 mL). The reaction was stirred at room temperature for 15 minutes andthen concentrated in vacuo. The residue was filtered and washed withcopious amounts of H₂O to yield 4.65 g (88%) compound 33 as a whitesolid. R_(f)=0.08 (5% MeOH/CH₂Cl₂).

MS(PI-FAB) 634, (M+H)⁺.

¹H NMR (CDCl₃) δ 9.09-9.08 (m, 1H), 8.87-8.85 (m, 1H), 8.16-8.14 (m,1H), 7.52-7.48 (m, 1H), 7.23-7.20 (d, 2H), 7.03-7.00 (d, 2H), 6.91-6.88(d, 1H), 4.78-4.70 (q, 1H), 4.59-4.46 (dd, 2H), 3.89 (s, 1H), 3.65-3.50(m, 4H), 3.09-3.06 (m, 2H), 2.92-2.88 (m, 4H), 1.43 (s, 9H), 1.19 (s,3H), 1.17 (s, 3H).

¹³C NMR (CDCl₃) δ 170.1, 167.9, 154.5, 153.9, 150.7, 148.8, 136.0,133.4, 133.2, 130.6, 124.1, 121.9, 83.0, 73.9, 55.0, 53.7, 50.7, 46.0,45.7, 45.0, 37.9, 29.3, 28.0, 24.0.Step 6: Preparation of compound 40

A 250 mL flask was charged with compound 33 (2.5 g, 3.9 mmol), CH₂Cl₂(40 mL), and sat. NaHCO₃ (40 mL). The reaction mixture was stirredvigorously at 0° C. for 15 minutes. Stirring was ceased and the layerswere allowed to separate. A 2.0 M solution of phosgene in toluene (7.9mL, 16 mmol) was quickly added to the reaction mixture which was stirredvigorously for 60 minutes maintaining the temperature at 0° C. Thelayers were separated and the aqueous phase was washed with CH₂Cl₂ (30mL). The combined organic layers were washed with 0.2 N citric acid,brine, dried over Na₂SO₄, filtered, and concentrated in vacuo to yield2.76 g (100%) white foam. The crude material was purified through asilica plug, eluting with 100% ethyl acetate, to yield 2.15 g (78%)compound 40 as a white foam. R_(f)=0.43 (3:1 ethyl acetate: hexanes).

¹H NMR (CDCl₃) δ 9.09-9.08 (m, 1H), 8.87-8.85 (m, 1H), 8.16-8.14 (d,1H), 7.52-7.48 (m, 1H), 7.25-7.22 (d, 2H), 7.03-7.01 (d, 2H), 6.90-6.88(d, 1H), 4.78-4.70 (q, 1H), 4.60-4.45 (dd, 2H), 3.88 (s, 1H), 3.79-3.65(m, 8H), 3.10-3.07 (m, 2H), 1.43 (s, 9H), 1.18 (s, 3H), 1.17 (s, 3H).

¹³C NMR (CDCl₃) δ 169.9, 167.9, 154.1, 153.6, 150.2, 148.5, 136.1,133.8, 130.6, 124.2, 121.7, 82.9, 73.7, 54.8, 53.8, 50.6, 48.3, 45.8,37.7, 29.2, 27.9, 23.9.Step 7: Preparation 2 kDa urea-linked mPEG conjugate t-butyl ester

The 2 kilodalton mPEG-amine (192 mg, 0.09 mmol) and DMAP (11 mg, 0.09mmol) were dissolved in CH₂Cl₂ (0.6 mL). Triethylamine (19.5 μL,0.14mmol) was added, followed by compound 40 (100 mg, 0.14 mmol). Thereaction mixture was heated to reflux for 20 hours. The reaction wasconcentrated in vacuo and the residue was taken up in MeOH (25 mL). 2%cross-linked polystyrene sulfonic acid resin (300 mg) was added andreaction vessel was swirled for 2 hours. The mixture was then filteredand concentrated in vacuo to yield 182 mg (˜50%) of a beige solid whichwas purified by HPLC Method B1 yielding 50.7 mg 2 kDa mPEG conjugatet-butyl ester as a white wax. R_(f)=0.12 (5% MeOH/CH₂Cl₂). HPLC MethodC1 determined conjugate to be >99% pure with no remaining compound 33 ormPEG-amine (retention time=1.924).

¹H NMR (CDCl₃) δ 8.21-8.18 (d, 1H), 7.23-7.21 (d, 2H), 7.03-7.00 (d,2H), 6.91-6.88 (d, 1H), 4.76-4.73 (q, 1H), 4.60-4.46 (dd, 2H), 3.91-3.86(m, 3H), 3.64 (bs, 184H), 3.37 (s, 3H), 3.09-3.06 (m, 3H), 1.43 (s, 9H),1.20 (s, 3H), 1.17 (s, 3H).Step 8: Preparation 2 kDa urea-linked mPEG conjugate carboxylic acid

The 2 kDa urea-linked mPEG conjugate t-butyl ester (94 mg, 0.04 mmol)was dissolved in formic acid (5 mL) and heated at 40° C. for 48 hours.The reaction was concentrated in vacuo to yield 88 mg (100%) beige gel,which was purified by HPLC Method A1 to yield 53.7 mg (-60%) of the freecarboxylic acid as a white wax. R_(f)=0.45 (7/3 MeOH:H₂O+0.1% TFA; C-18Reverse Phase). HPLC Method Cl determined conjugate to be >99% pure(retention time=2.188)

¹H NMR (CDCl₃) δ 9.07 (bs, 1H), 8.86-8.85 (m, 1H), 8.23-8.20 (d, 1H),7.59-7.55 (m, 1H), 7.26-7.21 (d, 2H), 7.02-6.96 (m, 2H), 4.82-4.80 (m,1H), 4.60-4.49-(dd, 2H), 3.99 (s, 1H), 3.62 (bs, 184H), 3.37 (s, 3H),3.15-3.13 (m, 2H), 1.25 (s, 3H), 1.23 (s, 3H).

Example 554 Preparation of 5 kDa urea-linked mPEG conjugate carboxylicacid

The 5 kDa urea-linked mPEG conjugate t-butyl ester was prepared in thesame manner as the 2 kDa conjugate above, using a 5 kDa mPEG-amine, andyielded 476 mg (˜90%) white solid. The crude material (200 mg, 0.04mmol) was deprotected in the same manner as above yielding 182 mg (100%)beige gum. This was purified by HPLC Method B1, yielding 74.5 mg of the5 kDa urea-linked mPEG conjugate carboxylic acid as a white powder.R_(f)=0.16 (7/3 MeOH:H₂O+0.1% TFA; C-18 Reverse Phase). HPLC Method C1determined conjugate to be >99% pure (retention time=2.260).

¹H NMR (CDCl₃) δ 9.07 (bs, 1H), 8.86-8.85 (m, 1H), 8.17-8.15 (d, 1H),7.54-7.50 (m, 1H), 7.26-7.22 (d, 2H), 7.03-7.00 (d, 2H), 6.95-6.93 .(d,1H), 5.46 (bs, 1H), 4.83-4.81 (m, 1H), 4.60-4.46 (dd, 2H), 3.93 (s, 1H),3.64 (bs, 490H), 3.37 (s, 3H), 3.16 (m, 3H), 1.22 (s, 6H).

Example 555 Preparation of 2 kDa carbamate-linked mPEG conjugate t-butylester

The carbamate linked conjugates were prepared based on a method modifiedfrom International Patent Publication Number WO 92/16555. Thus, a 2 kDamPEG-alcohol (500 mg, 0.25 mmol) was dried by azeotropic distillation intoluene (5 mL). The solution was cooled to room temperature and CH₂Cl₂(5 mL) was added, followed by a 2.0 M solution of phosgene in toluene(0.38 mL, 0.75 mrnol). The reaction was stirred at room temperature for18 hours and then concentrated in vacuo to yield 500 mg (100%) of the 2kDa mPEG chloroformate as a white solid. A solution of compound 33 (317mg, 0.5 mmol) in CH₂Cl₂ (3 mL) was added to the 2 kDa mPEG chloroformate(500 mg, 0.25 mmol) dissolved in CH₂Cl₂ (2 mL). Triethylamine (35 μl,0.25 mmol) was added and reaction was stirred at room temperature for 30minutes. The reaction mixture was concentrated in vacuo and the residuewas taken up in MeOH (10 mL). 2% cross-linked polystyrene sulfonic acidresin (750 mg) was added and the reaction vessel was swirled for 2hours. The mixture was then filtered and concentrated in vacuo to yield470 mg (75%) of the 2 kDa carbamate-linked mPEG conjugate t-butyl esteras a white solid. HPLC Method C1 shows >96% pure (retention time=2.639).

Example 556 Preparation of 2 kDa carbamate-linked mPEG conjugatecarboxylic acid

The crude 2 kDa carbamate-linked mPEG conjugate t-butyl ester (250 mg,0.1 mmol) was dissolved in formic acid (5 mL) and heated at 40° C. for48 hours. The reaction was concentrated in vacuo to yield 280 mg (190%)of the 2 kDa carbamate-linked mPEG conjugate carboxylic acid as a beigegel.

BIOLOGICAL EXAMPLES Example A In vitro Assay For Determining Binding ofCandidate Compounds to VLA-4

An in vitro assay was used to assess binding of candidate compounds toα₄β₁, integrin. Compounds which bind in this assay can be used to assessVCAM-1 levels in biological samples by conventional assays (e.g.,competitive binding assays). This assay is sensitive to IC₅₀ values aslow as about 1 nM.

The activity of α₄β₁ integrin was measured by the interaction of solubleVCAM-1 with Jurkat cells (e.g., American Type Culture Collection Nos.TIB 152, TIB 153, and CRL 8163), a human T-cell line which expresseshigh levels of α₄β₁ integrin. VCAM-1 interacts with the cell surface inan α₄β₁ integrin-dependent fashion (Yednock et al., J. Bio. Chem., 1995,270:28740).

Recombinant soluble VCAM-1 was expressed as a chimeric fuision proteincontaining the seven extracellular domains of VCAM-1 on the N-terminusand the human IgG₁ heavy chain constant region on the C-terminus. TheVCAM-1 fuision protein was made and purified by the manner described byYednock, supra.

Jurkat cells were grown in RPMI 1640 supplemented with 10% fetal bovineserum, penicillin, streptomycin and glutamine as described by Yednock,supra. Jurkat cells were incubated with 1.5 mM MnCl₂ and 5 μg/mL 15/7antibody for 30 minutes on ice. Mn⁺² activates the receptor to enhanceligand binding, and 15/7 is a monoclonal antibody that recognizes anactivated/ligand occupied conformation of α₄β₁ integrin and locks themolecule into this conformation thereby stabilizing the VCAM-1/α₄β₁integrin interaction. Yednock et al., supra. Antibodies similar to the15/7 antibody have been prepared by other investigators (Luque et al.,1996, J. Bio. Chem., 271:11067) and may be used in this assay.

Cells were then incubated for 30 minutes at room temperature withcandidate compounds, in various concentrations ranging from 66 μg/mL to0.01 μg/mL using a standard 5-point serial dilution. 15 μL solublerecombinant VCAM-1 fusion protein was then added to Jurkat cells andincubated for 30 minutes on ice. (Yednock et al., supra.). Cells werethen washed two times and resuspended in PE-conjugated goat F(ab′)₂anti-mouse IgG Fc (Immunotech, Westbrook, Me.) at 1:200 and incubated onice, in the dark, for 30 minutes. Cells were washed twice and analyzedwith a standard fluorescence activated cell sorter (“FACS”) analysis asdescribed in Yednock et al., supra.

Compounds having an IC₅₀ of less than about 15 μM possess bindingaffinity to α₄β₁.

When tested in this assay, the compounds (or the correspondingcarboxylic acids of the ester compounds, i.e., the prodrugs) have anIC₅₀ of 15 μM or less.

Example B Adjuvant-Induced Arthritis in Rats

Adjuvant induced arthritis (“AIA”) is an animal model useful in thestudy of rheumatoid arthritis (RA), which is induced by injecting M.tuberculosis in the base of the tail of Lewis rats. Between 10 and 15days following injection, animals develop a severe, progressivearthritis.

Generally, compounds are tested for their ability to alter hind pawswelling and bone damage resulting from adjuvant-induced edema in rats.To quantitate the inhibition of hind paw swelling resulting from AIA,two phases of inflammation have been defined: (1) the primary andsecondary injected hind paw, and (2) the secondary uninjected hind paw,which generally begins developing about eleven days from the inductionof inflammation in the injected paw. Reduction of the latter type ofinflammation is an indication of immunosuppressive activity. Cf. Chang,Arth. Rheum., 20, 1135-1141 (1977).

Using an animal model of RA, such as AIA, enables one to study thecellular events involved in the early stages of the disease. CD44expression on macrophages and lymphocytes is up-regulated during theearly development of adjuvant arthritis, whereas LFA-1 expression isup-regulated later in the development of the disease. Understanding theinteractions between adhesion molecules and endothelium at the earlieststages of adjuvant arthritis could lead to significant advances in themethods used in the treatment of RA.

Example C Anti-alpha 4-integrin Antibodies for Prophylactic,Semi-Therapeutic and Therapeutic Treatment

A murine collagen-induced arthritis (ClA) model was used to examine thepharmacological effect of anti-alpha4-integrin antibodiesprophylactically, semi-therapeutically and therapeutically. Dosing ofthe antibody was begun on day 20 for the prophylactic animals. Dosingfor the semi-therapeutic animals was started when 10% of the mice cohorthad a clinical score of 1. Therapeutic dosing started when the mice withClA had a clinical score of 1. The results seen with the treatment ofthe ClA mice is seen in FIG. 1. These results indicate that blockade ofthe alpha 4-integrin ameliorates disease severity and suggest thatalpha-4 integrins (VLA-4 and LPAM-1) play an important role in- ClA andmay constitute a valid target for RA.

Example D The Effect of Anti-Alpha 4, Anti-α₄β₇ and anti-VCAM-1Antibodies on ClA Animals

Peripheral blood leucocytes (PBLs) from mice treated with anti-alpha4,anti-α4β7 and anti-VCAM1 antibodies were collected at 4, 12 and 24 hoursafter antibody dosing. Cells were stained with goat anti-rat antibodiesto confirm antibodies in circulation. Anti-α₄β₇ and anti-VCAMIantibodies had minimal effect on disease severity in the murine ClAmodels (FIG. 2). In contrast, mice treated with anti-alpha-4 antibodyhad reduced severity of disease. These results demonstrated that VLA-4,not LPAM-1 or CAM-1 are involved in cell trafficking in ClA. Individualclinical scores on day 21 are shown in FIG. 3.

Example E Effect of the Compound of Formula P in Vivo in ClA AnimalModel

The effect of the compound of Formula P was evaluated in a therapeuticaClA model. It was demonstrated to have no effect on disease progressionin spite of average blood levels of 1 μg/mL at 16 hours after dosing.

100 DBA/1 LacJ male mice, ages 6-8 weeks were obtained from JacksonLaboratories. The mice were immunized at Day 0 as follows: 100 μg bovinetype II collagen (Chondrex) in Complete Freund's adjuvant (CFA, Sigma)were injected into the mice. At Day 21, another 100 μg bovine type IIcollagen (Chondrex) in Incomplete Freund's Adjuvant (IFA, Sigma) wasinjected into the mice. Treatment with the compound began when theanimals had the disease (i.e., 100% therapeutic). TABLE 12 Group nSubstance Dose 1 15 Untreated — 2 15 151246, 2% Tween/0/5% CMC 100 mg/kgb.i.d. 3 15 2% Tween/0.5% CMC 5 ml/kg b.i.d. 4 15 PS/2 (anti-VLA-4) 80μg/100 μL/mouse i.p. every 2 days 5 15 Control, Rag IgG2b 70 μg/100L/mouse i.p. every 2 daysThe mice are scored daily after the boost. When a mouse showed symptomsit was assigned to a treatment group (therapeutic protocol). Treatmentcontinued for 3-weeks-of dosing. Dosing was every other day for theantibody group and control group. FIG. 4 shows the effect of anti-VLA-4antibodies and the compound of Formula P on ClA animals.

Example F The Effect of the Compounds of Formulae W and Y in AIA Rats

It has been shown that anti-alpha 4 integrin antibody treatmentsignificantly reduced clinical and histological scores in an AIA model.The compound of Formula N at 30 mg/kg b.i.d. was shown to ameliorateclinical and histological scores in the AIA animal model. Thus, thecompounds of Formulae W and Y were analyzed at 30 mg/kg, 10 mg/kg and 3mg/kg amounts in ClA. The compound of Formula P was shown to amelioratedisease at all concentrations tested. PK results demonstrate that 12hours after dosing, the levels of the compound of Formula P were abovereceptor saturation at all conetrations tested. In contrast, thecompound of Formula N dose at 3 mg/kg had no effect on disease severity.Consistent with these results, the compound was present in the blood at12 hours was below receptor saturation.

The AIA animal models were Lewis rats (weight of 175-200 g) immunized onday 0 by injection in the base of the tail with 0.1 mg M. tuberculosisin mineral oil with 12 hour dosing. There were 6 rats per group in theefficacy study. Paw swelling and erythema (indicators of inducedcondition) were measured daily and Basal paws selected for histologicalanalysis at the end of the study. Plasma was collected at 4 and 12 hoursfor compound measurement. FIG. 5 shows the results obtained at thedifferent doses for each of the compounds tested. TABLE 13 Group DoseGroup Description Ab/vehicle Route Schedule Dose Volume 1 VehicleSaline, pH 4.1 s.c. b.i.d. n/a 5 mL/kg 2 Compound of Formula N Saline,pH 4.1 s.c. b.i.d. 30 mg/kg 5 mL/kg 3 Compound of Formula N Saline, pH4.1 s.c. b.i.d. 10 mg/kg 5 mL/kg 4 Compound of Formula N Saline, pH 4.1s.c. b.i.d. 3 mg/kg 5 mL/kg 5 Vehicle 2 2% Tween 80 + p.o. b.i.d. n/a 5mL/kg 0.5% CMC susp. 6 Compound of Formula P 2% Tween 80 + p.o. b.i.d.30 mg/kg 5 mL/kg 0.5% CMC susp. 7 Compound of Formula P 2% Tween 80 +p.o. b.i.d. 10 mg/kg 5 mL/kg 0.5% CMC susp. 8 Compound of Formula P 2%Tween 80 + p.o. b.i.d. 3 mg/kg 5 mL/kg 0.5% CMC susp.

Example G Evaluation of Compounds in ClA Animal Model

Anti alpha-4 antibodies, and the compounds of Formulae W and Y weretested in the rat ClA model according to the methods shown in Table 14below. TABLE 14 Dose Group Group Description AB/Vehicle Route ScheduleDose Volume 1 Murine IgG1 Mu anti-E. tenella i.v. Days (10-12 3 mg/kg 5mL/kg and 13-15) 2 Anti-VLA-4 Ab GG5/3 i.v. Days (10-12 3 mg/kg 5 mL/kgand 13-15) 3 Vehicle 1 Saline, pH 4.1 s.c. b.i.d. n/a 5 mL/kg 4 Compoundof Formula N Saline, pH 4.1 s.c. b.i.d. 30 mg/kg 5 mL/kg 5 Compound ofFormula N Saline, pH 4.1 s.c. b.i.d. 10 mg/kg 5 mL/kg 6 Compound ofFormula N Saline, pH 4.1 s.c. b.i.d. 3 mg/kg 5 mL/kg 7 Vehicle 2 2%Tween 80 + p.o. b.i.d. n/a 5 mL/kg 0.5% CMC susp. 8 Compound of FormulaP 2% Tween 80 + p.o. b.i.d. 30 mg/kg 5 mL/kg 0.5% CMC susp. 9 Compoundof Formula P 2% Tween 80 + p.o. b.i.d. 10 mg/kg 5 mL/kg 0.5% CMC susp.10 Compound of Formula P 2% Tween 80 + p.o. b.i.d. 3 mg/kg 5 mL/kg 0.5%CMC susp.Results indicated that anti-alpha-4 antibody treatment amelioratesdisease. However, neither the compound of Formula N nor Y had any impacton disease severity. At 12 hours, animals administered the compound ofFormula P had blood levels of the compound that were above receptorsaturation at all concentrations tests.

The results in FIG. 6 show the results for compounds. There were 8 ratsper group in the efficacy study. Paw swelling and erythema were measureddaily and the tarsal paws collected for histological analysis at the endof the study. Plasma which was heparinized was collected at 12 hours(i.e., the trough level) and at 4 horus after the last dose for compoundmeasurement (FIG. 6).

Example H Prophylactic Treatment of ClA Animals

In order to examine the role of VLA-4 in RA, two models have beenevaluated: murine ClA (collagen-induced arthritis) and rat AIA(adjuvant-induced arthritis). Initial studies evaluated the effect ofanti-α4-integrin antibodies on disease progression in both models.

Prophylactic treatment with anti-α 4-integrin antibodies (PS/2) in theClA model resulted in a 50% reduction of clinical scores (see FIG. 7 andTable 15 below). -The results strongly suggest the VLA-4 pathway mayplay a role in ClA, but was somewhat offset the SD among mice/group wasgreater than desired. DBA/1 LacJ mice, aged 8 weeks will be immunizedwith bovine collagen (100 μg/mice) in CFA and re-challenge on day 21 inIFA. TABLE 15 Group Description Antibody Dose n 1 Disease control Nonen/a 10 2 Anti-VLA-4 mAb PS/2 (IgG2b) 80 μg/mouse, 10 3×/week 3 Rat IgRat anti-human 80 μg/mouse, 10 creatinine 3×/week kinase (IgG1) 4Vehicle control, 100 μL/mouse, 10 PBS 3×/weekAntibody treatment started on day 20 and ended on day 42. Clinicalevaluation: paw swelling. Where “n” is the number of animals in thegroup.

Example I Therapeutic Treatment of AIA Animals with Anti-α4-integrinAntibody

Therapeutic treatment with anti-α4-integrin antibody (GG5/3) in the AIAmodel resulted in significant reduction ofjoint and histological scores(FIGS. 8 and 9). These results indicate that in-this model, blockade ofα4-integrin significantly reduces disease severity, synovitis and mankinscores. No isotype antibody control was included in this experiment.

Lewis rats (animal weight is about 175 to about 200 g) were immunized onday 0 by injection in the base of the tail with 0.1 mg M. tuberculosisin mineral oil. Injections were performed as follows: TABLE 16 GroupDescription Antibody Dose N 1 Disease control None N/a 5 2 Anti-VLA-4mAb GG5/3 (IgG1), 3 mg/kg 5 I.V. days 8, 11, 14 and 17N represents the number of animals in the group.

Clinical evaluation was perforrnzed by measuring paw erythema andswelling. Histological evaluation was performed by staining tarsaljoints with hematoxylin/eosin and saffranin o-fast green stain.

Example J Exposure, Potency and Specificity of Compounds in AIA Model

The compound of Tanabe is included as a comparative example and forms apositive control. The compound is depicted by E. Kudlacz et al.,Pharmacol. Exp. Ther., 301(2):747-52 (2002) and below:

The goal of these studies was to evaluate the efficacy of selected VLA-4small molecule antagonist in the AIA model. Compounds with differentcharacteristics related to exposure (the compound of Formula M versusthe compound of Formula N), potency (for example, the compound ofFormula N versus the compound of Formula P) and/or specificity (forexample, the compound of Formula N versus the compound of Tanabe) wereassessed for their effect on disease.

Results indicate that the compounds of Formulae V and W ameloriatedisease at the administered dose.

Lewis rats (animal weight is about 175 to about 200 g) were immunized onday 0 by injection in the base of the tail with 0.1 mg M. tuberculosisin mineral oil. Six rats (“n”) per group for clinical and histologicalevaluation; 3 rats per group for clinical evaluation and “tissuedistribution” determinations. Plasma collected twelve hr (trough) andfour hr after last dosing for PK. Animals were dosed as shown in Table17 below. The results are displayed in FIG. 10. TABLE 17 GroupDescription Ab/vehicle Route Schedule Dose Dose Voume Efficacy Exposure1 Muring IgG1 Mu anti-E. tenella i.v. Days 8, 11, 14 3 mg/kg 5 ml/kg N =6 N/A 2 Anti-VLA-4 Ab GG5/3 i.v. Days 8, 11, 14 3 mg/kg 5 ml/kg N = 6N/A 3 Vehicle 1 Saline, pH 4.1 s.c. b.i.d.* N/A 5 ml/kg N = 6 N/A 4Compound of Saline, pH 4.1 s.c. b.i.d.* 30 mg/kg 5 mg/kg N = 6 N = 3Formula N 5 Vehicle 2 10% EtOH:90% corn oil s.c. b.i.d.* N/A 5 mg/kg N =6 N = 3 6 Compound of 10% EtOH:90% corn oil s.c. b.i.d.* 100 mg/kg 5mg/kg N = 6 N = 3 Formula N

Example K Potency and Specificity of Compounds in the AIA Model

The results indicate that all compounds tested ameloriate diseasescores. The results are displayed in FIG. 11. The compounds of FormulaeW and Y affected disease equally well in spite of their differentpotencies. Compounds with preferential selectivity for α4β1 ameloriatedisease better than those with equal selectivity for α4β1 and α4β7.

Lewis rats (animal weight was about 175 to about 200 g) were immunizedon day 0 by injection in the base of the tail with 0.1 mg M.tuberculosis in mineral oil. Six rats per group were evaluated forclinical and histological scores. Plasma was collected four hours afterlast dosing for PK. TABLE 18 Dose Group Description Ab/vehicle RouteSchedule Dose volume Efficacy 1 Vehicle Saline, pH 4.1 s.c. b.i.d. n/a 5mg/kg N = 6 2 Compound of Formula N Saline, pH 4.1 s.c. b.i.d.  30mg/kg  5 mg/kg N = 6 3 Vehicle 2 20% Tween 80 + 0.5% CMC susp. p.o.b.i.d. n/a 5 mg/kg N = 6 4 Compound of Formula P 20% Tween 80 + 0.5% CMCsusp. p.o. b.i.d.  30 mg/kg  5 mg/kg N = 6 5 Vehicle 3 NaOH/dH₂O p.o.b.i.d. n/a 5 mg/kg N = 6 6 Compound of Formula O NaOH/dH₂O p.o. b.i.d.100 mg/kg  5 mg/kg N = 6 7 Compound of Tanabe NaOH/dH₂O s.c. b.i.d.  30mg/kg  5 mg/kg N = 6

While the present invention has been described with reference to thespecific embodiments thereof, it should be understood by those skilledin the art that various changes may be made and equivalents may besubstituted without departing from the true spirit and scope of theinvention. In addition, many modifications may be made to adapt aparticular situation, material, composition of matter, process, processstep or steps, to the objective, spirit and scope of the presentinvention. All such modifications are intended to be within the scope ofthe invention.

All references cited herein are herein incorporated by reference intheir entirety for all purposes.

1. A method for treating rheumatoid arthritis in a subject in needthereof comprising administering a combination therapy comprisingmethotrexate and an antibody to alpha-4 integrin or an immunologicallyactive antigen binding fragment thereof to the subject, intherapeutically effective amounts.
 2. The method of claim 1, wherein thesubject is a mammal.
 3. The method of claim 2, wherein the mammal is ahuman.
 4. The method of claim 1, wherein the antibody or theimmunologically active antigen binding fragment thereof is a monoclonalantibody or an immunologically active fragment of a monoclonal antibody.5. The method of claim 1, wherein the antibody or immunologically activeantigen binding fragment thereof binds to alpha-4 integrin such that itinhibits binding to VCAM-1 or inhibit α4β1 dimer activity.
 6. The methodof claim 1, wherein the antibody is a humanized antibody or a humanizedimmunologically active antigen binding fragment thereof.
 7. The methodof claim 6, wherein the humanized antibody is natalizumab or animmunologically active fragment thereof.
 8. The method of claim 7,wherein natalizumab is administered intravenously or subcutaneously. 9.The method of any of claims 1-8, wherein the immunologically activeantigen binding fragment of the antibody is Fab, scFv, or F(ab′)₂. 10.The method of claim 8, wherein the natalizumab is administeredsubcutaneously in a dosage of about 0.01 mg/kg of body weight to about50 mg/kg of body weight.
 11. The method of any of claims 1-8, whereinthe antibody or immunologically active antigen binding fragment thereofare administered in a series of doses separated by intervals of days orweeks.
 12. The method of any of claim 1-8, wherein an additionalanti-inflammatory composition is also administered to the mammal, intherapeutically effective amounts.
 13. The method according to claim 1,wherein the combination therapy is administered in a series of dosesseparated by intervals of days or weeks.
 14. The method of claim 1,wherein the immunolobulin, when administered to a subject in needthereof, reaches a blood level of immunoglobulin in the subject of about10 ng/ml or more.
 15. The method of claim 1, wherein the the antibody isadministered via injection at a dose of about 2.0 mg/kg to about 8.0mg/kg dosage.
 16. The method of claim 1, wherein the combination therapyfurther comprises an adjuvant.
 17. The method of any of claims 1-8,wherein the methotrexate is administered in a dose of about 2 mg toabout 20 mg.
 18. A regimen for the treatment of rheumatoid arthritiswhich comprises administering to a subject in need thereof about 2 mg toabout 20 mg of methotrexate and about 0.01 mg/kg of body weight to about50 mg/kg of body weight of an antibody to alpha-4 integrin or animmunologically active antigen binding fragment thereof, wherein theamount of methotrexate administered per week does not exceed 20 mg.